Novel anti-baff antibodies

ABSTRACT

The present invention relates to anti-BAFF antibody molecules, including novel humanized anti-BAFF antibodies, therapeutic and diagnostic methods and compositions for using the same.

SEQUENCE LISTING

The instant application contains a Sequence Listing which has beensubmitted electronically in ASCII format and is hereby incorporated byreference in its entirety. Said ASCII copy, created on Jan. 30, 2015, isnamed 09-0625-US-2_SL.txt and is 256,600 bytes in size.

TECHNICAL FIELD OF THE INVENTION

This invention generally relates to anti-BAFF antibodies for diagnosticand therapeutic use. More specifically, anti-BAFF antibodies and methodsfor treating various diseases or disorders are disclosed. Pharmaceuticalcompositions and kits comprising such compounds are also disclosed.

BACKGROUND OF THE INVENTION

B-cell activating factor (BAFF) is a cytokine that belongs to the tumornecrosis factor (TNF) ligand superfamily and acts as a ligand forreceptors BAFF-R (BR3), TACI (transmembrane activator and calciummodulator and cyclophilin ligand interactor) and BCMA (B-cell maturationantigen). The interaction between BAFF and its receptors triggerssignals essential for the formation and maintenance of B cells, which inturn synthesizes immunoglobulins in response to invasion by a foreignsubstance. Appropriate levels of BAFF in a patient help maintain normallevels of immunity whereas inadequate levels can lead toimmunodeficiency and excessive levels can result in abnormally highantibody production.

When a patient exhibits autoimmunity, it produces antibodies against thetissues or organs of its own body. Autoimmune diseases, including lupuserythematosus and rheumatoid arthritis, result from excessive levels ofBAFF in the body. Thus it is important to modulate the production ofBAFF in order to treat the patients having these diseases.

BAFF can exist in three forms: membrane bound (mbBAFF), soluble trimericBAFF (sBAFF) and a multimeric form consisting of 60 BAFF monomers. Therelative importance of the various forms of BAFF in normal and diseasephysiology is not well understood. As noted, BAFF binds to threereceptors, BAFFR (BR3), TACI and BCMA. A proliferation-inducing ligand(APRIL), a related member of the TNF receptor ligand family, has beenshown to bind with high affinity to TACI and BCMA. In contrast to thehigh affinity APRIL:BCMA interaction, the BAFF:BCMA interaction is oflow affinity (1-2 μM) and is not believed to be play an important rolein vivo (Bossen and Schneider, 2006).

Soluble BAFF is expressed at high levels in individuals with systemiclupus erythematosus (SLE) and in inflamed target organs such as thekidney. Soluble BAFF serves as a critical factor for B cell homeostasisand survival (Kalled et al., 2005; Mackay et al., 2003; Smith andCancro, 2003; Patke et al., 2004). Autoantibody formation byBAFF-dependent B cells results in glomerular IC deposits, initially atthe glomerular basement membrane (GBM), mesangium and interstitialtissue within the proximal tubular epithelial cells (PTEC). These ICdeposits lead to complement fixation and neutrophil activation resultingin local kidney damage. Inflammatory mediators (e.g. IL6, IL8, MCP-1)produced by the damaged kidney cells (MC, PTEC, renal fibroblasts,endothelial cells) fuel an inflammatory cycle by increasing immune cellinfiltration (e.g. B cells, T cells, dendritic cells, neutrophils andmacrophages).

Anti-BAFF monoclonal antibody belimumab (Benlysta®) has demonstratedactivity in the treatment of systemic lupus erythematosus (SLE) and hasthe demonstrated ability to decrease autoantibody formation. Belimumabis currently approved for the treatment of active SLE without kidneyinvolvement. Belimumab, however, is not reported to bind to mbBAFF butinhibition of sBAFF only is therefore a viable path to treat excessivelevels of BAFF and increased antibody production. In contrast, theanti-BAFF peptibody blisibimod (A-623) and the anti-BAFF mAb tabalumab(LY2127399) have been reported to bind both sBAFF and mbBAFF (2010Anthera press release and 2012 Lilly press release). Given the uncertainroles for various forms of BAFF in disease, antagonist molecules againstsBAFF and mbBAFF with beneficial pharmacologic properties may possessadded benefit in the treatment of immunological and autoimmune diseasesin humans.

BRIEF SUMMARY OF THE INVENTION

The present invention relates to novel anti-BAFF antibodies fortreatment of immunological and autoimmune diseases, including, but notlimited to, systemic lupus erythematosus, lupus nephritis and rheumatoidarthritis. The anti-BAFF antibodies of this invention bind to human BAFFwith high affinity thus inhibiting abnormally high immunoglobulinproduction. In one embodiment of the invention anti-BAFF antibodies arederived from mouse hybridomas, for example monoclonal antibodies.Another embodiment includes full length anti-BAFF antibodies. In yetanother embodiment, the present invention provides anti-BAFF humanantibodies, including full-length humanized monoclonal anti-BAFFantibodies. Further embodiments encompass DNA molecules encodingantibodies of the present invention, expression vectors and host cellscomprising such DNA molecules, and methods of making antibodies of thepresent invention. The present invention further provides therapeuticuses for the antibodies of the present invention, in particularimmunological and autoimmune diseases.

In one embodiment, the invention provides an anti-BAFF antibody moleculecomprising a light chain variable domain with a CDR1 selected from thegroup consisting of SEQ ID NO: 1, SEQ ID NO: 5, SEQ ID NO: 10, SEQ IDNO: 13, SEQ ID NO: 15, SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78, SEQID NO: 79, SEQ ID NO: 80, SEQ ID NO: 249, SEQ ID NO: 250, SEQ ID NO:251, SEQ ID NO: 252, SEQ ID NO: 253, SEQ ID NO: 254, SEQ ID NO: 255, SEQID NO: 256, SEQ ID NO: 257, SEQ ID NO: 258, SEQ ID NO: 259, SEQ ID NO:260, SEQ ID NO: 261, SEQ ID NO: 262, SEQ ID NO: 263, SEQ ID NO: 264, SEQID NO: 265, SEQ ID NO: 266, SEQ ID NO: 267, SEQ ID NO: 268, SEQ ID NO:269, SEQ ID NO: 270, SEQ ID NO: 271, SEQ ID NO: 272, SEQ ID NO: 273, SEQID NO: 274 and SEQ ID NO: 275; a CDR2 selected from the group consistingof SEQ ID NO: 2, SEQ ID NO: 6, SEQ ID NO: 8, SEQ ID NO: 11, SEQ ID NO:16, SEQ ID NO: 276, SEQ ID NO: 277, SEQ ID NO: 278, SEQ ID NO: 279, SEQID NO: 280, SEQ ID NO: 281, SEQ ID NO: 282, SEQ ID NO: 283, SEQ ID NO:284, SEQ ID NO: 285, SEQ ID NO: 286, SEQ ID NO: 287, SEQ ID NO: 288, SEQID NO: 289, SEQ ID NO: 290, SEQ ID NO: 291 and SEQ ID NO: 292; and aCDR3 selected from the group consisting of SEQ ID NO: 3, SEQ ID NO: 4,SEQ ID NO: 7, SEQ ID NO: 9, SEQ ID NO: 12, SEQ ID NO:14, SEQ ID NO: 17,SEQ ID NO: 293, SEQ ID NO: 294, SEQ ID NO: 295, SEQ ID NO: 296, SEQ IDNO: 297, SEQ ID NO: 298, SEQ ID NO: 299, SEQ ID NO: 300, SEQ ID NO: 301,SEQ ID NO: 302, SEQ ID NO: 303, SEQ ID NO: 304, SEQ ID NO: 305, SEQ IDNO: 306, SEQ ID NO: 307, SEQ ID NO: 308, SEQ ID NO: 309, SEQ ID NO: 310and SEQ ID NO: 311; and a heavy chain variable domain with a CDR1selected from the group consisting of SEQ ID NO: 18, SEQ ID NO: 21, SEQID NO: 23, SEQ ID NO: 25, SEQ ID NO: 28, SEQ ID NO: 31, SEQ ID NO: 34,SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 81, SEQ ID NO: 312, SEQ ID NO:313, SEQ ID NO: 314, SEQ ID NO: 315, SEQ ID NO: 316, SEQ ID NO: 317, SEQID NO: 318, SEQ ID NO: 319, SEQ ID NO: 320, SEQ ID NO: 321, SEQ ID NO:322, SEQ ID NO: 323, SEQ ID NO: 324, SEQ ID NO: 325, SEQ ID NO: 326, SEQID NO: 327, SEQ ID NO: 328, SEQ ID NO: 329, SEQ ID NO: 330, SEQ ID NO:331, SEQ ID NO: 332, SEQ ID NO: 392, SEQ ID NO: 333, SEQ ID NO: 334, SEQID NO: 335 and SEQ ID NO: 336; a CDR2 selected from the group consistingof SEQ ID NO: 19, SEQ ID NO: 24, SEQ ID NO: 26, SEQ ID NO: 29, SEQ IDNO: 32, SEQ ID NO: 35, SEQ ID NO: 38, SEQ ID NO: 337, SEQ ID NO: 338,SEQ ID NO: 339, SEQ ID NO: 340, SEQ ID NO: 341, SEQ ID NO: 342, SEQ IDNO: 343, SEQ ID NO: 344, SEQ ID NO: 343, SEQ ID NO: 345, SEQ ID NO: 346,SEQ ID NO: 347, SEQ ID NO: 348, SEQ ID NO: 349, SEQ ID NO: 350, SEQ IDNO: 351, SEQ ID NO: 352, SEQ ID NO: 353, SEQ ID NO: 354, SEQ ID NO: 355,SEQ ID NO: 356, SEQ ID NO: 357, SEQ ID NO: 358, SEQ ID NO: 359, SEQ IDNO: 360, SEQ ID NO: 361, SEQ ID NO: 362, SEQ ID NO: 363, SEQ ID NO: 364,SEQ ID NO: 365, SEQ ID NO: 366 and SEQ ID NO: 367; and a CDR3 selectedfrom the group consisting of SEQ ID NO: 20, SEQ ID NO: 22, SEQ ID NO:27, SEQ ID NO: 30, SEQ ID NO: 33, SEQ ID NO. 39, SEQ ID NO: 368, SEQ IDNO: 369, SEQ ID NO: 370, SEQ ID NO: 371, SEQ ID NO: 372, SEQ ID NO: 373,SEQ ID NO: 374, SEQ ID NO: 375, SEQ ID NO: 376, SEQ ID NO: 377, SEQ IDNO: 378, SEQ ID NO: 378, SEQ ID NO: 379, SEQ ID NO:380, SEQ ID NO: 381,SEQ ID NO: 382, SEQ ID NO: 383, SEQ ID NO: 384, SEQ ID NO: 385, SEQ IDNO: 386, SEQ ID NO: 387, SEQ ID NO: 388, SEQ ID NO: 389, SEQ ID NO: 390and SEQ ID NO: 391.

In other embodiments, the invention provides (a) an anti-BAFF antibodymolecule where the light chain variable domain comprises a CDR1 of SEQID NO: 1, a CDR2 of SEQ ID NO: 2 and a CDR3 of SEQ ID NO: 3 and a heavychain variable domain comprises a CDR1 of SEQ ID NO: 18, a CDR2 of SEQID NO: 19 and a CDR3 of SEQ ID NO: 20; (b) an anti-BAFF antibodymolecule where the light chain variable domain comprises a CDR1 of SEQID NO: 1, a CDR2 of SEQ ID NO: 2 and a CDR3 of SEQ ID NO: 4 and a heavychain variable domain comprises a CDR1 of SEQ ID NO: 21, a CDR2 of SEQID NO: 19 and a CDR3 of SEQ ID NO: 22; (c) an anti-BAFF antibodymolecule where the light chain variable domain comprises a CDR1 of SEQID NO: 1, a CDR2 of SEQ ID NO: 2 and a CDR3 of SEQ ID NO: 4 and a heavychain variable domain comprises a CDR1 of SEQ ID NO: 23, a CDR2 of SEQID NO: 24 and a CDR3 of SEQ ID NO: 20; (d) an anti-BAFF antibodymolecule where the light chain variable domain comprises a CDR1 of SEQID NO: 5, a CDR2 of SEQ ID NO: 6 and a CDR3 of SEQ ID NO: 7 and a heavychain variable domain comprises a CDR1 of SEQ ID NO: 25, a CDR2 of SEQID NO: 26 and a CDR3 of SEQ ID NO: 27; (e) an anti-BAFF antibodymolecule where the light chain variable domain comprises a CDR1 of SEQID NO: 5, a CDR2 of SEQ ID NO: 8 and a CDR3 of SEQ ID NO: 9 and a heavychain variable domain comprises a CDR1 of SEQ ID NO: 28, a CDR2 of SEQID NO: 29 and a CDR3 of SEQ ID NO: 30; (f) an anti-BAFF antibodymolecule where the light chain variable domain comprises a CDR1 of SEQID NO: 10, a CDR2 of SEQ ID NO: 11 and a CDR3 of SEQ ID NO: 12 and aheavy chain variable domain comprises a CDR1 of SEQ ID NO: 31, a CDR2 ofSEQ ID NO: 32 and a CDR3 of SEQ ID NO: 33; (g) an anti-BAFF antibodymolecule where the light chain variable domain comprises a CDR1 of SEQID NO: 13, a CDR2 of SEQ ID NO: 6 and a CDR3 of SEQ ID NO: 14 and aheavy chain variable domain comprises a CDR1 of SEQ ID NO: 34, a CDR2 ofSEQ ID NO: 35 and a CDR3 of SEQ ID NO: 27; (h) an anti-BAFF antibodymolecule where the light chain variable domain comprises a CDR1 of SEQID NO: 10, a CDR2 of SEQ ID NO: 6 and a CDR3 of SEQ ID NO: 7 and a heavychain variable domain comprises a CDR1 of SEQ ID NO: 36, a CDR2 of SEQID NO: 26 and a CDR3 of SEQ ID NO: 27; (i) an anti-BAFF antibodymolecule where the light chain variable domain comprises a CDR1 of SEQID NO: 15, a CDR2 of SEQ ID NO: 16 and a CDR3 of SEQ ID NO: 17 and aheavy chain variable domain comprises a CDR1 of SEQ ID NO: 37, a CDR2 ofSEQ ID NO: 38 and a CDR3 of SEQ ID NO: 39; (j) an anti-BAFF antibodymolecule where the light chain variable domain comprises a CDR1 of SEQID NO: 76, a CDR2 of SEQ ID NO: 16 and a CDR3 of SEQ ID NO: 17 and aheavy chain variable domain comprises a CDR1 of SEQ ID NO: 37, a CDR2 ofSEQ ID NO: 38 and a CDR3 of SEQ ID NO: 39; (k) an anti-BAFF antibodymolecule where the light chain variable domain comprises a CDR1 of SEQID NO: 77, a CDR2 of SEQ ID NO: 16 and a CDR3 of SEQ ID NO: 17 and aheavy chain variable domain comprises a CDR1 of SEQ ID NO: 37, a CDR2 ofSEQ ID NO: 38 and a CDR3 of SEQ ID NO: 39; (l) an anti-BAFF antibodymolecule where the light chain variable domain comprises a CDR1 of SEQID NO: 78, a CDR2 of SEQ ID NO: 16 and a CDR3 of SEQ ID NO: 17 and aheavy chain variable domain comprises a CDR1 of SEQ ID NO: 37, a CDR2 ofSEQ ID NO: 38 and a CDR3 of SEQ ID NO: 39; (m) an anti-BAFF antibodymolecule where the light chain variable domain comprises a CDR1 of SEQID NO: 79, a CDR2 of SEQ ID NO: 16 and a CDR3 of SEQ ID NO: 17 and aheavy chain variable domain comprises a CDR1 of SEQ ID NO: 37, a CDR2 ofSEQ ID NO: 38 and a CDR3 of SEQ ID NO: 39; (n) an anti-BAFF antibodymolecule where the light chain variable domain comprises a CDR1 of SEQID NO: 80, a CDR2 of SEQ ID NO: 16 and a CDR3 of SEQ ID NO: 17 and aheavy chain variable domain comprises a CDR1 of SEQ ID NO: 37, a CDR2 ofSEQ ID NO: 38 and a CDR3 of SEQ ID NO: 39; (o) an anti-BAFF antibodymolecule where the light chain variable domain comprises a CDR1 of SEQID NO: 5, a CDR2 of SEQ ID NO: 8 and a CDR3 of SEQ ID NO: 9 and a heavychain variable domain comprises a CDR1 of SEQ ID NO: 81, a CDR2 of SEQID NO: 29 and a CDR3 of SEQ ID NO: 30; (p) an anti-BAFF antibodymolecule where the light chain variable domain comprises a CDR1 of SEQID NO: 249, a CDR2 of SEQ ID NO: 276 and a CDR3 of SEQ ID NO: 293 and aheavy chain variable domain comprises a CDR1 of SEQ ID NO: 312, a CDR2of SEQ ID NO: 337 and a CDR3 of SEQ ID NO: 368; (q) an anti-BAFFantibody molecule where the light chain variable domain comprises a CDR1of SEQ ID NO: 250, a CDR2 of SEQ ID NO: 276 and a CDR3 of SEQ ID NO: 293and a heavy chain variable domain comprises a CDR1 of SEQ ID NO: 312, aCDR2 of SEQ ID NO: 337 and a CDR3 of SEQ ID NO: 368; (r) an anti-BAFFantibody molecule where the light chain variable domain comprises a CDR1of SEQ ID NO: 251, a CDR2 of SEQ ID NO: 277 and a CDR3 of SEQ ID NO: 294and a heavy chain variable domain comprises a CDR1 of SEQ ID NO: 313, aCDR2 of SEQ ID NO: 338 and a CDR3 of SEQ ID NO: 369; (s) an anti-BAFFantibody molecule where the light chain variable domain comprises a CDR1of SEQ ID NO: 252, a CDR2 of SEQ ID NO: 278 and a CDR3 of SEQ ID NO: 295and a heavy chain variable domain comprises a CDR1 of SEQ ID NO: 314, aCDR2 of SEQ ID NO: 339 and a CDR3 of SEQ ID NO: 370; (t) an anti-BAFFantibody molecule where the light chain variable domain comprises a CDR1of SEQ ID NO: 253, a CDR2 of SEQ ID NO: 279 and a CDR3 of SEQ ID NO: 296and a heavy chain variable domain comprises a CDR1 of SEQ ID NO: 315, aCDR2 of SEQ ID NO: 340 and a CDR3 of SEQ ID NO: 371; (u) an anti-BAFFantibody molecule where the light chain variable domain comprises a CDR1of SEQ ID NO: 15, a CDR2 of SEQ ID NO: 16 and a CDR3 of SEQ ID NO: 297and a heavy chain variable domain comprises a CDR1 of SEQ ID NO: 316, aCDR2 of SEQ ID NO: 341 and a CDR3 of SEQ ID NO: 39; (v) an anti-BAFFantibody molecule where the light chain variable domain comprises a CDR1of SEQ ID NO: 254, a CDR2 of SEQ ID NO: 280 and a CDR3 of SEQ ID NO: 298and a heavy chain variable domain comprises a CDR1 of SEQ ID NO: 317, aCDR2 of SEQ ID NO: 342 and a CDR3 of SEQ ID NO: 372; (w) an anti-BAFFantibody molecule where the light chain variable domain comprises a CDR1of SEQ ID NO: 255, a CDR2 of SEQ ID NO: 281 and a CDR3 of SEQ ID NO: 298and a heavy chain variable domain comprises a CDR1 of SEQ ID NO: 317, aCDR2 of SEQ ID NO: 343 and a CDR3 of SEQ ID NO: 373; (x) an anti-BAFFantibody molecule where the light chain variable domain comprises a CDR1of SEQ ID NO: 256, a CDR2 of SEQ ID NO: 2 and a CDR3 of SEQ ID NO: 299and a heavy chain variable domain comprises a CDR1 of SEQ ID NO: 317, aCDR2 of SEQ ID NO: 344 and a CDR3 of SEQ ID NO: 372; (y) an anti-BAFFantibody molecule where the light chain variable domain comprises a CDR1of SEQ ID NO: 255, a CDR2 of SEQ ID NO: 281 and a CDR3 of SEQ ID NO: 298and a heavy chain variable domain comprises a CDR1 of SEQ ID NO: 318, aCDR2 of SEQ ID NO: 343 and a CDR3 of SEQ ID NO: 374; (z) an anti-BAFFantibody molecule where the light chain variable domain comprises a CDR1of SEQ ID NO: 257, a CDR2 of SEQ ID NO: 282 and a CDR3 of SEQ ID NO: 300and a heavy chain variable domain comprises a CDR1 of SEQ ID NO: 319, aCDR2 of SEQ ID NO: 345 and a CDR3 of SEQ ID NO: 375; (aa) an anti-BAFFantibody molecule where the light chain variable domain comprises a CDR1of SEQ ID NO: 258, a CDR2 of SEQ ID NO: 283 and a CDR3 of SEQ ID NO: 301and a heavy chain variable domain comprises a CDR1 of SEQ ID NO: 320, aCDR2 of SEQ ID NO: 346 and a CDR3 of SEQ ID NO: 376; (bb) an anti-BAFFantibody molecule where the light chain variable domain comprises a CDR1of SEQ ID NO: 259, a CDR2 of SEQ ID NO: 281 and a CDR3 of SEQ ID NO: 298and a heavy chain variable domain comprises a CDR1 of SEQ ID NO: 317, aCDR2 of SEQ ID NO: 347 and a CDR3 of SEQ ID NO: 377; (cc) an anti-BAFFantibody molecule where the light chain variable domain comprises a CDR1of SEQ ID NO: 260, a CDR2 of SEQ ID NO: 284 and a CDR3 of SEQ ID NO: 294and a heavy chain variable domain comprises a CDR1 of SEQ ID NO: 321, aCDR2 of SEQ ID NO: 348 and a CDR3 of SEQ ID NO: 378; (dd) an anti-BAFFantibody molecule where the light chain variable domain comprises a CDR1of SEQ ID NO: 254, a CDR2 of SEQ ID NO: 2 and a CDR3 of SEQ ID NO: 299and a heavy chain variable domain comprises a CDR1 of SEQ ID NO: 322, aCDR2 of SEQ ID NO: 349 and a CDR3 of SEQ ID NO: 372; (ee) an anti-BAFFantibody molecule where the light chain variable domain comprises a CDR1of SEQ ID NO: 261, a CDR2 of SEQ ID NO: 285 and a CDR3 of SEQ ID NO: 294and a heavy chain variable domain comprises a CDR1 of SEQ ID NO: 323, aCDR2 of SEQ ID NO: 350 and a CDR3 of SEQ ID NO: 378; (ff) an anti-BAFFantibody molecule where the light chain variable domain comprises a CDR1of SEQ ID NO: 262, a CDR2 of SEQ ID NO: 286 and a CDR3 of SEQ ID NO: 302and a heavy chain variable domain comprises a CDR1 of SEQ ID NO: 324, aCDR2 of SEQ ID NO: 351 and a CDR3 of SEQ ID NO: 379; (gg) an anti-BAFFantibody molecule where the light chain variable domain comprises a CDR1of SEQ ID NO: 263, a CDR2 of SEQ ID NO: 6 and a CDR3 of SEQ ID NO: 303and a heavy chain variable domain comprises a CDR1 of SEQ ID NO: 317, aCDR2 of SEQ ID NO: 352 and a CDR3 of SEQ ID NO: 380; (hh) an anti-BAFFantibody molecule where the light chain variable domain comprises a CDR1of SEQ ID NO: 264, a CDR2 of SEQ ID NO: 276 and a CDR3 of SEQ ID NO: 304and a heavy chain variable domain comprises a CDR1 of SEQ ID NO: 325, aCDR2 of SEQ ID NO: 353 and a CDR3 of SEQ ID NO: 381; (ii) an anti-BAFFantibody molecule where the light chain variable domain comprises a CDR1of SEQ ID NO: 265, a CDR2 of SEQ ID NO: 287 and a CDR3 of SEQ ID NO: 305and a heavy chain variable domain comprises a CDR1 of SEQ ID NO: 326, aCDR2 of SEQ ID NO: 354 and a CDR3 of SEQ ID NO: 382; (jj) an anti-BAFFantibody molecule where the light chain variable domain comprises a CDR1of SEQ ID NO: 266, a CDR2 of SEQ ID NO: 287 and a CDR3 of SEQ ID NO: 306and a heavy chain variable domain comprises a CDR1 of SEQ ID NO: 326, aCDR2 of SEQ ID NO: 355 and a CDR3 of SEQ ID NO: 383; (kk) an anti-BAFFantibody molecule where the light chain variable domain comprises a CDR1of SEQ ID NO: 267, a CDR2 of SEQ ID NO: 285 and a CDR3 of SEQ ID NO: 294and a heavy chain variable domain comprises a CDR1 of SEQ ID NO: 327, aCDR2 of SEQ ID NO: 356 and a CDR3 of SEQ ID NO: 369; (ll) an anti-BAFFantibody molecule where the light chain variable domain comprises a CDR1of SEQ ID NO: 268, a CDR2 of SEQ ID NO: 276 and a CDR3 of SEQ ID NO: 306and a heavy chain variable domain comprises a CDR1 of SEQ ID NO: 328, aCDR2 of SEQ ID NO: 357 and a CDR3 of SEQ ID NO: 383; (mm) an anti-BAFFantibody molecule where the light chain variable domain comprises a CDR1of SEQ ID NO: 269, a CDR2 of SEQ ID NO: 288 and a CDR3 of SEQ ID NO: 304and a heavy chain variable domain comprises a CDR1 of SEQ ID NO: 329, aCDR2 of SEQ ID NO: 358 and a CDR3 of SEQ ID NO: 384; (nn) an anti-BAFFantibody molecule where the light chain variable domain comprises a CDR1of SEQ ID NO: 270, a CDR2 of SEQ ID NO: 276 and a CDR3 of SEQ ID NO: 306and a heavy chain variable domain comprises a CDR1 of SEQ ID NO: 330, aCDR2 of SEQ ID NO: 359 and a CDR3 of SEQ ID NO: 385; (oo) an anti-BAFFantibody molecule where the light chain variable domain comprises a CDR1of SEQ ID NO: 271, a CDR2 of SEQ ID NO: 289 and a CDR3 of SEQ ID NO: 307and a heavy chain variable domain comprises a CDR1 of SEQ ID NO: 331, aCDR2 of SEQ ID NO: 360 and a CDR3 of SEQ ID NO: 385; (pp) an anti-BAFFantibody molecule where the light chain variable domain comprises a CDR1of SEQ ID NO: 261, a CDR2 of SEQ ID NO: 285 and a CDR3 of SEQ ID NO: 294and a heavy chain variable domain comprises a CDR1 of SEQ ID NO: 332, aCDR2 of SEQ ID NO: 361 and a CDR3 of SEQ ID NO: 386; (qq) an anti-BAFFantibody molecule where the light chain variable domain comprises a CDR1of SEQ ID NO: 272, a CDR2 of SEQ ID NO: 289 and a CDR3 of SEQ ID NO: 307and a heavy chain variable domain comprises a CDR1 of SEQ ID NO: 331, aCDR2 of SEQ ID NO: 362 and a CDR3 of SEQ ID NO: 385; (rr) an anti-BAFFantibody molecule where the light chain variable domain comprises a CDR1of SEQ ID NO: 266, a CDR2 of SEQ ID NO: 287 and a CDR3 of SEQ ID NO: 306and a heavy chain variable domain comprises a CDR1 of SEQ ID NO: 326, aCDR2 of SEQ ID NO: 355 and a CDR3 of SEQ ID NO: 383; (ss) an anti-BAFFantibody molecule where the light chain variable domain comprises a CDR1of SEQ ID NO: 270, a CDR2 of SEQ ID NO: 276 and a CDR3 of SEQ ID NO: 306and a heavy chain variable domain comprises a CDR1 of SEQ ID NO: 330, aCDR2 of SEQ ID NO: 359 and a CDR3 of SEQ ID NO: 285; (tt) an anti-BAFFantibody molecule where the light chain variable domain comprises a CDR1of SEQ ID NO: 270, a CDR2 of SEQ ID NO: 276 and a CDR3 of SEQ ID NO: 306and a heavy chain variable domain comprises a CDR1 of SEQ ID NO: 392, aCDR2 of SEQ ID NO: 363 and a CDR3 of SEQ ID NO: 387; (uu) an anti-BAFFantibody molecule where the light chain variable domain comprises a CDR1of SEQ ID NO: 273, a CDR2 of SEQ ID NO: 276 and a CDR3 of SEQ ID NO: 308and a heavy chain variable domain comprises a CDR1 of SEQ ID NO: 333, aCDR2 of SEQ ID NO: 364 and a CDR3 of SEQ ID NO: 388; (vv) an anti-BAFFantibody molecule where the light chain variable domain comprises a CDR1of SEQ ID NO: 274, a CDR2 of SEQ ID NO: 290 and a CDR3 of SEQ ID NO: 309and a heavy chain variable domain comprises a CDR1 of SEQ ID NO: 334, aCDR2 of SEQ ID NO: 365 and a CDR3 of SEQ ID NO: 389; (ww) an anti-BAFFantibody molecule where the light chain variable domain comprises a CDR1of SEQ ID NO: 275, a CDR2 of SEQ ID NO: 291 and a CDR3 of SEQ ID NO: 310and a heavy chain variable domain comprises a CDR1 of SEQ ID NO: 335, aCDR2 of SEQ ID NO: 366 and a CDR3 of SEQ ID NO: 390; and (xx) ananti-BAFF antibody molecule where the light chain variable domaincomprises a CDR1 of SEQ ID NO: 258, a CDR2 of SEQ ID NO: 292 and a CDR3of SEQ ID NO: 311 and a heavy chain variable domain comprises a CDR1 ofSEQ ID NO: 336, a CDR2 of SEQ ID NO: 367 and a CDR3 of SEQ ID NO: 391.

In another embodiment of the present invention, the anti-BAFF antibodymolecule comprises a light chain variable region of any one of SEQ IDNOS: 82-97, and a heavy chain variable region of any one of SEQ ID NOS:100-115. In a preferred embodiment, the present invention providesmonoclonal antibodies with the combinations of light chain variable andheavy chain variable regions of SEQ ID NO: 82/101, 88/101, 94/112 or93/114.

In an additional embodiment of the present invention, the anti-BAFFantibody molecule neutralizes all three forms of human BAFF, the formsof which include membrane bound (mbBAFF), soluble trimeric BAFF, andsoluble 60-mer BAFF. In particular, the anti-BAFF antibody molecules ofthe present invention neutralize human soluble 60-mer BAFF. Furthermore,the anti-BAFF antibody molecules of the present invention neutralizehuman soluble trimeric BAFF. Finally, the anti-BAFF antibody moleculesof the present invention neutralize human membrane-bound BAFF.

In a further embodiment, the present invention relates to an anti-BAFFantibody molecule comprising a humanized light chain variable domaincomprising the CDRs of SEQ ID NO: 76, 16 and 17 and framework regionshaving an amino acid sequence at least 90% identical, at least 93%identical or at least 95% identical to the amino acid sequence of theframework regions of the variable domain light chain amino acid sequenceof SEQ ID NO: 82 and a humanized heavy chain variable domain comprisingthe CDRs of SEQ ID NO: 37, 38 and 39 and framework regions having anamino acid sequence at least 90% identical, at least 93% identical or atleast 95% identical to the amino acid sequence of the framework regionsof the variable domain heavy chain amino acid sequence of SEQ ID NO:101. In one embodiment, the anti-BAFF antibody molecule is a humanizedmonoclonal antibody.

In a further embodiment, the present invention relates to an anti-BAFFantibody molecule comprising a humanized light chain variable domaincomprising the CDRs of SEQ ID NO: 15, 16 and 17 and framework regionshaving an amino acid sequence at least 90% identical, at least 93%identical or at least 95% identical to the amino acid sequence of theframework regions of the variable domain light chain amino acid sequenceof SEQ ID NO: 88 and a humanized heavy chain variable domain comprisingthe CDRs of SEQ ID NO: 37, 38 and 39 and framework regions having anamino acid sequence at least 90% identical, at least 93% identical or atleast 95% identical to the amino acid sequence of the framework regionsof the variable domain heavy chain amino acid sequence of SEQ ID NO:101. In one embodiment, the anti-BAFF antibody molecule is a humanizedmonoclonal antibody.

In a further embodiment, the present invention relates to an anti-BAFFantibody molecule comprising a humanized light chain variable domaincomprising the CDRs of SEQ ID NO:5, 8 and 9 and framework regions havingan amino acid sequence at least 90% identical, at least 93% identical orat least 95% identical to the amino acid sequence of the frameworkregions of the variable domain light chain amino acid sequence of SEQ IDNO: 94 and a humanized heavy chain variable domain comprising the CDRsof SEQ ID NO: 81, 29 and 30 and framework regions having an amino acidsequence at least 90% identical, at least 93% identical or at least 95%identical to the amino acid sequence of the framework regions of thevariable domain heavy chain amino acid sequence of SEQ ID NO: 112. Inone embodiment, the anti-BAFF antibody molecule is a humanizedmonoclonal antibody.

In a further embodiment, the present invention relates to an anti-BAFFantibody molecule comprising a humanized light chain variable domaincomprising the CDRs of SEQ ID NO: 5, 8 and 9 and framework regionshaving an amino acid sequence at least 90% identical, at least 93%identical or at least 95% identical to the amino acid sequence of theframework regions of the variable domain light chain amino acid sequenceof SEQ ID NO: 93 and a humanized heavy chain variable domain comprisingthe CDRs of SEQ ID NO: 81, 29 and 30 and framework regions having anamino acid sequence at least 90% identical, at least 93% identical or atleast 95% identical to the amino acid sequence of the framework regionsof the variable domain heavy chain amino acid sequence of SEQ ID NO:114. In one embodiment, the anti-BAFF antibody molecule is a humanizedmonoclonal antibody.

In yet another embodiment, the anti-BAFF antibody molecule is amonoclonal antibody or a humanized monoclonal antibody.

The present invention also provides pharmaceutical compositionscomprising an anti-BAFF antibody molecule described herein and apharmaceutically acceptable carrier.

The present invention further provides a method for treating a subjecthaving a BAFF-associated disorder comprising administering to thesubject an anti-BAFF antibody molecule, or a pharmaceutical compositioncomprising an anti-BAFF antibody molecule and a pharmaceuticallyacceptable carrier, which anti-BAFF antibody molecule binds to humanBAFF. Specifically provided in the present invention is a method fortreating an inflammatory disease, an autoimmune disease, a respiratorydisease, a metabolic disorder or cancer comprising administering to asubject in need thereof an effective amount of an anti-BAFF antibodymolecule, or a pharmaceutical composition comprising an anti-BAFFantibody molecule and a pharmaceutically acceptable carrier. Inparticular, the disease to be treated may be systemic lupuserythematosus, lupus nephritis or rheumatoid arthritis.

Also provided in the invention is a method for inhibiting the binding ofBAFF to one or more BAFF receptors on a mammalian cell, wherein the BAFFreceptor is BAFF-R (BR3), TACI (transmembrane activator and calciummodulator and cyclophilin ligand interactor) and/or BCMA (B-cellmaturation antigen), comprising administering to the cell an anti-BAFFantibody molecule, whereby signaling mediated by the BAFF receptor isinhibited.

A further embodiment encompasses a DNA molecule encoding a variablelight chain region, variable heavy chain region, light chain region orheavy chain region described herein.

In one embodiment, an isolated polynucleotide comprises a sequenceencoding a light chain variable region of any one of SEQ ID NOS: 82-97,or a heavy chain variable region of any one of SEQ ID NOS: 100-115. Inanother embodiment, the isolated polynucleotide comprises a light chainvariable region is SEQ ID NO: 234 and the heavy chain variable region isSEQ ID NO: 396, the light chain variable region is SEQ ID NO: 393 andthe heavy chain variable region is SEQ ID NO: 396, the light chainvariable region is SEQ ID NO: 395 and the heavy chain variable region isSEQ ID NO: 397 or the light chain variable region is SEQ ID NO: 394 andthe heavy chain variable region is SEQ ID NO: 398.

Another embodiment encompasses an expression vector containing a DNAmolecule. A further embodiment encompasses a host cell carrying one ormore expression vectors. In one embodiment, a host is a mammalian cell.

A further embodiment encompasses a method for producing an antibodymolecule comprising transfecting a mammalian host cell with one or morevectors, cultivating the host cell and recovering and purifying theantibody molecule.

Another aspect of the invention relates to a method for producing anantibody molecule comprising obtaining a mammalian host cell comprisingone or more of the vectors above, and cultivating the host cell. In oneembodiment, the method further comprises recovering and purifying theantibody molecule.

In one embodiment, the present invention further provides an antibodymolecule above for use in medicine. In one embodiment, the use is thetreatment of an inflammatory disease, of an autoimmune disease, of arespiratory disease, of a metabolic disorder or of cancer. In oneembodiment, the use is for the treatment of systemic lupuserythematosus, lupus nephritis or rheumatoid arthritis. In anotherembodiment, the use of the antibody molecule is for the preparation of amedicament for the treatment of an inflammatory disease, of anautoimmune disease, of a respiratory disease, of a metabolic disorder orof cancer, preferably for the treatment of systemic lupus erythematosus,lupus nephritis or rheumatoid arthritis. In yet another embodiment, thepresent invention provides a method for inhibiting the binding of BAFFto one or more BAFF receptors on a mammalian cell that is not within ahuman, wherein the method comprises contacting the mammalian cell withan antibody molecule according to the present invention.

BRIEF DESCRIPTION OF DRAWINGS

FIG. 1: Anti-BAFF monoclonal antibody potency against sBAFF: chimericHuIgG1 KO vs. parental mouse monoclonal antibodies.

FIG. 2: Anti-BAFF monoclonal antibody against mbBAFF: chimeric HuIgG1 KOvs. parental mouse monoclonal antibodies.

DETAILED DESCRIPTION

The present invention provides antibodies that bind to BAFF, inparticular human BAFF. The present invention also relates to humanizedantibodies. In specific embodiments, the sequence of these humanizedantibodies has been identified based on the sequences of certain leadmouse antibodies.

The lead mouse antibodies of the present invention were derived frommouse hybridomas. The immunization of the mice is carried out usingdifferent techniques. For example, antibodies that are specific forhuman BAFF can be raised against an immunogenic antigen such as anisolated BAFF protein, and/or a portion thereof of any of the above(including synthetic peptides). Preparation of immunogenic antigens andmonoclonal antibody production can be performed using any suitabletechnique known in the art.

The lead mouse antibodies were selected based on their high affinity toBAFF. Accordingly, in one aspect, the present invention provides anantibody that binds to human BAFF with high affinity. Selected mouseantibodies were humanized to result in humanized antibodies. Thehumanized antibodies of the present invention bind to human BAFF withhigh affinity. Accordingly, in another aspect, the present inventionprovides a humanized antibody that binds to human BAFF with highaffinity.

Accordingly, in one embodiment, the present invention provides ananti-BAFF antibody having a K_(D) of less than 100 pM. In a furtherembodiment, the present invention provides an anti-BAFF antibody havinga K_(D) of less than 10 pM. In a further embodiment, the presentinvention provides an anti-BAFF antibody having a K_(D) less than 1 pM.

In a further aspect, a humanized monoclonal anti-BAFF antibody of thepresent invention has favorable biophysical properties, for examplequality, stability, or solubility.

In one aspect, the anti-BAFF antibody is a humanized antibody. In oneaspect, the anti-BAFF antibody is a monoclonal antibody. In one aspect,the anti-BAFF antibody is a full length antibody. In one aspect, theanti-BAFF antibody is a humanized monoclonal antibody, for example afull length humanized monoclonal antibody.

An anti-BAFF antibody of the present invention recognizes a specific or“BAFF epitope”. Epitopes may be determined by various techniques knownin the art, such as X-ray crystallography, Hydrogen/Deuterium ExchangeMass Spectrometry (HXMS), site-directed mutagenesis, alanine scanningmutagenesis, and peptide screening methods.

DEFINITIONS

The generalized structure of antibodies or immunoglobulin is well knownto those of skill in the art. These molecules are heterotetramericglycoproteins, typically of about 150,000 daltons, composed of twoidentical light (L) chains and two identical heavy (H) chains and aretypically referred to as full length antibodies. Each light chain iscovalently linked to a heavy chain by one disulfide bond to form aheterodimer, and the heterotrameric molecule is formed through acovalent disulfide linkage between the two identical heavy chains of theheterodimers. Although the light and heavy chains are linked together byone disulfide bond, the number of disulfide linkages between the twoheavy chains varies by immunoglobulin isotype. Each heavy and lightchain also has regularly spaced intrachain disulfide bridges. Each heavychain has at the amino-terminus a variable domain (V_(H)), followed bythree or four constant domains (C_(H1), C_(H2), C_(H3), and C_(H4)), aswell as a hinge region between C_(H1) and C_(H2). Each light chain hastwo domains, an amino-terminal variable domain (V_(L)) and acarboxy-terminal constant domain (C_(L)). The V_(L) domain associatesnon-covalently with the V_(H) domain, whereas the C_(L) domain iscommonly covalently linked to the C_(H1) domain via a disulfide bond.Particular amino acid residues are believed to form an interface betweenthe light and heavy chain variable domains (Chothia et al., 1985, J.Mol. Biol. 186:651-663). Variable domains are also referred herein asvariable regions.

Certain domains within the variable domains differ extensively betweendifferent antibodies i.e., are “hypervariable.” These hypervariabledomains contain residues that are directly involved in the binding andspecificity of each particular antibody for its specific antigenicdeterminant. Hypervariability, both in the light chain and the heavychain variable domains, is concentrated in three segments known ascomplementarity determining regions (CDRs) or hypervariable loops(HVLs). CDRs are defined by sequence comparison in Kabat et al., 1991,In: Sequences of Proteins of Immunological Interest, 5^(th) Ed. PublicHealth Service, National Institutes of Health, Bethesda, Md., whereasHVLs (also referred herein as CDRs) are structurally defined accordingto the three-dimensional structure of the variable domain, as describedby Chothia and Lesk, 1987, J. Mol. Biol. 196: 901-917. These two methodsresult in slightly different identifications of a CDR. As defined byKabat, CDR-L1 is positioned at about residues 24-34, CDR-L2, at aboutresidues 50-56, and CDR-L3, at about residues 89-97 in the light chainvariable domain; CDR-H1 is positioned at about residues 31-35, CDR-H2 atabout residues 50-65, and CDR-H3 at about residues 95-102 in the heavychain variable domain. The exact residue numbers that encompass aparticular CDR will vary depending on the sequence and size of the CDR.Those skilled in the art can routinely determine which residues comprisea particular CDR given the variable region amino acid sequence of theantibody. The CDR1, CDR2, CDR3 of the heavy and light chains thereforedefine the unique and functional properties specific for a givenantibody.

The three CDRs within each of the heavy and light chains are separatedby framework regions (FR), which contain sequences that tend to be lessvariable. From the amino terminus to the carboxy terminus of the heavyand light chain variable domains, the FRs and CDRs are arranged in theorder: FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4. The largely β-sheetconfiguration of the FRs brings the CDRs within each of the chains intoclose proximity to each other as well as to the CDRs from the otherchain. The resulting conformation contributes to the antigen bindingsite (see Kabat et al., 1991, NIH Publ. No. 91-3242, Vol. I, pages647-669), although not all CDR residues are necessarily directlyinvolved in antigen binding.

FR residues and Ig constant domains are not directly involved in antigenbinding, but contribute to antigen binding and/or mediate antibodyeffector function. Some FR residues are thought to have a significanteffect on antigen binding in at least three ways: by noncovalentlybinding directly to an epitope, by interacting with one or more CDRresidues, and by affecting the interface between the heavy and lightchains. The constant domains are not directly involved in antigenbinding but mediate various Ig effector functions, such as participationof the antibody in antibody dependent cellular cytotoxicity (ADCC),antibody dependent cellular phagocytosis (ADCP) and complement dependentcytotoxicity (CDC).

The light chains of vertebrate immunoglobulins are assigned to one oftwo clearly distinct classes, kappa (κ) and lambda (λ), based on theamino acid sequence of the constant domain. By comparison, the heavychains of mammalian immunoglobulins are assigned to one of five majorclasses, according to the sequence of the constant domains: IgA, IgD,IgE, IgG, and IgM. IgG and IgA are further divided into subclasses(isotypes), e.g., IgG₁, IgG₂, IgG₃, IgG₄, IgA₁, and IgA₂. The heavychain constant domains that correspond to the different classes ofimmunoglobulins are called α, β, ε, γ and μ, respectively. The subunitstructures and three-dimensional configurations of the classes of nativeimmunoglobulins are well known.

The terms, “antibody”, “anti-BAFF antibody”, “anti-BAFF antibodymolecule”, “humanized anti-BAFF antibody”, “humanized anti-BAFF epitopeantibody”, and “variant humanized anti-BAFF epitope antibody”specifically encompass monoclonal antibodies (including full lengthmonoclonal antibodies), polyclonal antibodies, multispecific antibodies,and antibody fragments such as variable domains and other portions ofantibodies that exhibit a desired biological activity, e.g., BAFFbinding. The term “monoclonal antibody” (mAb) refers to an antibody thatis highly specific, being directed against a single antigenicdeterminant, an “epitope”. Therefore, the modifier “monoclonal” isindicative of antibodies directed to the identical epitope and is not tobe construed as requiring production of the antibody by any particularmethod. It should be understood that monoclonal antibodies can be madeby any technique or methodology known in the art; including e.g., thehybridoma method (Kohler et al., 1975, Nature 256:495), or recombinantDNA methods known in the art (see, e.g., U.S. Pat. No. 4,816,567), ormethods of isolation of monoclonal recombinantly produced using phageantibody libraries, using techniques described in Clackson et al., 1991,Nature 352: 624-628, and Marks et al., 1991, J. Mol. Biol. 222: 581-597.

The term “monomer” refers to a homogenous form of an antibody. Forexample, for a full-length antibody, monomer means a monomeric antibodyhaving two identical heavy chains and two identical light chains.

Chimeric antibodies consist of the heavy and light chain variableregions of an antibody from one species (e.g., a non-human mammal suchas a mouse) and the heavy and light chain constant regions of anotherspecies (e.g., human) antibody and can be obtained by linking the DNAsequences encoding the variable regions of the antibody from the firstspecies (e.g., mouse) to the DNA sequences for the constant regions ofthe antibody from the second (e.g. human) species and transforming ahost with an expression vector containing the linked sequences to allowit to produce a chimeric antibody. Alternatively, the chimeric antibodyalso could be one in which one or more regions or domains of the heavyand/or light chain is identical with, homologous to, or a variant of thecorresponding sequence in a monoclonal antibody from anotherimmunoglobulin class or isotype, or from a consensus or germlinesequence. Chimeric antibodies can include fragments of such antibodies,provided that the antibody fragment exhibits the desired biologicalactivity of its parent antibody, for example binding to the same epitope(see, e.g., U.S. Pat. No. 4,816,567; and Morrison et al., 1984, Proc.Natl. Acad. Sci. USA 81: 6851-6855).

The terms “antibody fragment”, “anti-BAFF antibody fragment”, “anti-BAFFantibody molecule”, “anti-BAFF epitope antibody fragment”, “humanizedanti-BAFF antibody fragment”, “humanized anti-BAFF epitope antibodyfragment”, “variant humanized anti-BAFF epitope antibody fragment” referto a portion of a full length anti-BAFF antibody, in which a variableregion or a functional capability is retained, for example, specificBAFF epitope binding. Examples of antibody fragments include, but arenot limited to, a Fab, Fab′, F(ab′)₂, Fd, Fv, scFv and scFv-Fc fragment,a diabody, a linear antibody, a single-chain antibody, a minibody, adiabody formed from antibody fragments, and multispecific antibodiesformed from antibody fragments.

Full length antibodies can be treated with enzymes such as papain orpepsin to generate useful antibody fragments. Papain digestion is usedto produces two identical antigen-binding antibody fragments called“Fab” fragments, each with a single antigen-binding site, and a residual“Fc” fragment. The Fab fragment also contains the constant domain of thelight chain and the C_(H1) domain of the heavy chain. Pepsin treatmentyields a F(ab′)₂ fragment that has two antigen-binding sites and isstill capable of cross-linking antigen.

Fab′ fragments differ from Fab fragments by the presence of additionalresidues including one or more cysteines from the antibody hinge regionat the C-terminus of the C_(H1) domain. F(ab′)₂ antibody fragments arepairs of Fab′ fragments linked by cysteine residues in the hinge region.Other chemical couplings of antibody fragments are also known.

“Fv” fragment contains a complete antigen-recognition and binding siteconsisting of a dimer of one heavy and one light chain variable domainin tight, non-covalent association. In this configuration, the threeCDRs of each variable domain interact to define an antigen-biding siteon the surface of the V_(H)-V_(L) dimer. Collectively, the six CDRsconfer antigen-binding specificity to the antibody.

A “single-chain Fv” or “scFv” antibody fragment is a single chain Fvvariant comprising the V_(H) and V_(L) domains of an antibody where thedomains are present in a single polypeptide chain. The single chain Fvis capable of recognizing and binding antigen. The scFv polypeptide mayoptionally also contain a polypeptide linker positioned between theV_(H) and V_(L) domains in order to facilitate formation of a desiredthree-dimensional structure for antigen binding by the scFv (see, e.g.,Pluckthun, 1994, In The Pharmacology of monoclonal Antibodies, Vol. 113,Rosenburg and Moore eds., Springer-Verlag, New York, pp. 269-315).

A “diabody” refers to small antibody fragments with two antigen-bindingsites, which fragments comprise a heavy chain variable domain (V_(H))connected to a light chain variable domain (V_(L)) in the samepolypeptide chain (V_(H)-V_(L) or V_(L)-V_(H)). Diabodies are describedmore fully in, e.g., Holliger et al. (1993) Proc. Natl. Acad. Sci. USA90: 6444-6448.

Other recognized antibody fragments include those that comprise a pairof tandem Fd segments (V_(H)-C_(H1)-V_(H)-C_(H1)) to form a pair ofantigen binding regions. These “linear antibodies” can be bispecific ormonospecific as described in, for example, Zapata et al. 1995, ProteinEng. 8(10):1057-1062.

A “humanized antibody” or a “humanized antibody fragment” is a specifictype of chimeric antibody which includes an immunoglobulin amino acidsequence variant, or fragment thereof, which is capable of binding to apredetermined antigen and which, comprises one or more FRs havingsubstantially the amino acid sequence of a human immunoglobulin and oneor more CDRs having substantially the amino acid sequence of a non-humanimmunoglobulin. This non-human amino acid sequence often referred to asan “import” sequence is typically taken from an “import” antibodydomain, particularly a variable domain. In general, a humanized antibodyincludes at least the CDRs or HVLs of a non-human antibody, insertedbetween the FRs of a human heavy or light chain variable domain. Thepresent invention describes specific humanized BAFF antibodies whichcontain CDRs derived from the mouse monoclonal antibodies or humanizedCDRs shown in Tables 3 and 4 inserted between the FRs of human germlinesequence heavy and light chain variable domains. It will be understoodthat certain mouse FR residues may be important to the function of thehumanized antibodies and therefore certain of the human germlinesequence heavy and light chain variable domains residues are modified tobe the same as those of the corresponding mouse sequence.

In another aspect, a humanized BAFF antibody comprises substantially allof at least one, and typically two, variable domains (such as contained,for example, in Fab, Fab′, F(ab′)2, Fabc, and Fv fragments) in whichall, or substantially all, of the CDRs correspond to those of anon-human immunoglobulin, and specifically herein, all of the CDRs aremouse or humanized sequences as detailed in Tables 1 through 4 hereinbelow and all, or substantially all, of the FRs are those of a humanimmunoglobulin consensus or germline sequence. In another aspect, ahumanized anti-BAFF antibody also includes at least a portion of animmunoglobulin Fc region, typically that of a human immunoglobulin.Ordinarily, the antibody will contain both the light chain as well as atleast the variable domain of a heavy chain. The antibody also mayinclude one or more of the C_(H1), hinge, C_(H2), C_(H3), and/or C_(H4)regions of the heavy chain, as appropriate.

A humanized anti-BAFF antibody can be selected from any class ofimmunoglobulins, including IgM, IgG, IgD, IgA and IgE, and any isotype,including IgG₁, IgG₂, IgG₃, IgG₄, IgA₁ and IgA₂. For example, theconstant domain can be a complement fixing constant domain where it isdesired that the humanized antibody exhibit cytotoxic activity, and theisotype is typically IgG₁. Where such cytotoxic activity is notdesirable, the constant domain may be of another isotype, e.g., IgG₂. Analternative humanized anti-BAFF antibody can comprise sequences frommore than one immunoglobulin class or isotype, and selecting particularconstant domains to optimize desired effector functions is within theordinary skill in the art. In specific embodiments, the presentinvention provides antibodies that are IgG1 antibodies and moreparticularly, are IgG1 antibodies in which there is a knock-out ofeffector functions.

The FRs and CDRs, or HVLs, of a humanized anti-BAFF antibody need notcorrespond precisely to the parental sequences. For example, one or moreresidues in the import CDR, or HVL, or the consensus or germline FRsequence may be altered (e.g., mutagenized) by substitution, insertionor deletion such that the resulting amino acid residue is no longeridentical to the original residue in the corresponding position ineither parental sequence but the antibody nevertheless retains thefunction of binding to BAFF. Such alteration typically will not beextensive and will be conservative alterations. Usually, at least 75% ofthe humanized antibody residues will correspond to those of the parentalconsensus or germline FR and import CDR sequences, more often at least90%, and most frequently greater than 95%, or greater than 98% orgreater than 99%.

Immunoglobulin residues that affect the interface between heavy andlight chain variable regions (“the V_(L)-V_(H) interface”) are thosethat affect the proximity or orientation of the two chains with respectto one another. Certain residues that may be involved in interchaininteractions include V_(L) residues 34, 36, 38, 44, 46, 87, 89, 91, 96,and 98 and V_(H) residues 35, 37, 39, 45, 47, 91, 93, 95, 100, and 103(utilizing the numbering system set forth in Kabat et al., Sequences ofProteins of Immunological Interest (National Institutes of Health,Bethesda, Md., 1987)). U.S. Pat. No. 6,407,213 also discusses thatresidues such as V_(L) residues 43 and 85, and V_(H) residues 43 and 60also may be involved in this interaction. While these residues areindicated for human IgG only, they are applicable across species.Important antibody residues that are reasonably expected to be involvedin interchain interactions are selected for substitution into theconsensus sequence.

The terms “consensus sequence” and “consensus antibody” refer to anamino acid sequence which comprises the most frequently occurring aminoacid residue at each location in all immunoglobulins of any particularclass, isotype, or subunit structure, e.g., a human immunoglobulinvariable domain. The consensus sequence may be based on immunoglobulinsof a particular species or of many species. A “consensus” sequence,structure, or antibody is understood to encompass a consensus humansequence as described in certain embodiments, and to refer to an aminoacid sequence which comprises the most frequently occurring amino acidresidues at each location in all human immunoglobulins of any particularclass, isotype, or subunit structure. Thus, the consensus sequencecontains an amino acid sequence having at each position an amino acidthat is present in one or more known immunoglobulins, but which may notexactly duplicate the entire amino acid sequence of any singleimmunoglobulin. The variable region consensus sequence is not obtainedfrom any naturally produced antibody or immunoglobulin. Kabat et al.,1991, Sequences of Proteins of Immunological Interest, 5th Ed. PublicHealth Service, National Institutes of Health, Bethesda, Md., andvariants thereof. The FRs of heavy and light chain consensus sequences,and variants thereof, provide useful sequences for the preparation ofhumanized anti-BAFF antibodies. See, for example, U.S. Pat. Nos.6,037,454 and 6,054,297.

Human germline sequences are found naturally in the human population. Acombination of those germline genes generates antibody diversity.Germline antibody sequences for the light chain of the antibody comefrom conserved human germline kappa or lambda v-genes and j-genes.Similarly the heavy chain sequences come from germline v-, d- andj-genes (LeFranc, M-P, and LeFranc, G, “The Immunoglobulin Facts Book”Academic Press, 2001).

As used herein, “variant”, “anti-BAFF variant”, “humanized anti-BAFFvariant”, or “variant humanized anti-BAFF” each refers to a humanizedanti-BAFF antibody having at least a light chain variable murine CDRfrom any of the sequences as shown in Table 1 or a heavy chain murineCDR sequence derived from the murine monoclonal antibody as shown inTable 2. Variants include those having one or more amino acid changes inone or both light chain or heavy chain variable domains, provided thatthe amino acid change does not substantially impair binding of theantibody to BAFF.

An “isolated” antibody is one that has been identified and separatedand/or recovered from a component of its natural environment.Contaminant components of the antibody's natural environment are thosematerials that may interfere with diagnostic or therapeutic uses of theantibody, and can be enzymes, hormones, or other proteinaceous ornonproteinaceous solutes. In one aspect, the antibody will be purifiedto at least greater than 95% isolation by weight of antibody.

An isolated antibody includes an antibody in situ within recombinantcells in which it is produced, since at least one component of theantibody's natural environment will not be present. Ordinarily however,an isolated antibody will be prepared by at least one purification stepin which the recombinant cellular material is removed.

An “antibody molecule” refers to any one of the antibody definitionsdescribed above or an antigen-binding fragment thereof.

The term “antibody performance” refers to factors that contribute toantibody recognition of antigen or the effectiveness of an antibody invivo. Changes in the amino acid sequence of an antibody can affectantibody properties such as folding, and can influence physical factorssuch as initial rate of antibody binding to antigen (k_(a)),dissociation constant of the antibody from antigen (k_(d)), affinityconstant of the antibody for the antigen (Kd), conformation of theantibody, protein stability, and half life of the antibody.

The term “neutralize” generally relates to rendering inactive viainhibition of bioactivity. The term “inhibition” generally relates to asituation where a molecule is unable to execute its function. Inchemistry or biology the term “inhibit” means to limit, prevent or blockthe action or function, i.e., to inhibit an enzyme, or to inhibit achemical reaction. IC50 represents the concentration of a drug that isrequired for 50% inhibition in vitro and IC 90 represents theconcentration of a drug that is required for 90% inhibition in vitro.

The term “epitope tagged” when used herein, refers to an anti-BAFFantibody fused to an “epitope tag”. An “epitope tag” is a polypeptidehaving a sufficient number of amino acids to provide an epitope forantibody production, yet is designed such that it does not interferewith the desired activity of the humanized anti-BAFF antibody. Theepitope tag is usually sufficiently unique such that an antibody raisedagainst the epitope tag does not substantially cross-react with otherepitopes. Suitable tag polypeptides generally contain at least 6 aminoacid residues and usually contain about 8 to 50 amino acid residues, orabout 9 to 30 residues. Examples of epitope tags and the antibody thatbinds the epitope include the flu HA tag polypeptide and its antibody12CA5 (Field et al., 1988 Mol. Cell. Biol. 8: 2159-2165; c-myc tag and8F9, 3C7, 6E10, G4, B7 and 9E10 antibodies thereto (Evan et al., 1985,Mol. Cell. Biol. 5(12):3610-3616; and Herpes simplex virus glycoproteinD (gD) tag and its antibody (Paborsky et al. 1990, Protein Engineering3(6): 547-553). In certain embodiments, the epitope tag is a “salvagereceptor binding epitope”. As used herein, the term “salvage receptorbinding epitope” refers to an epitope of the Fc region of an IgGmolecule (such as IgG₁, IgG₂, IgG₃, or IgG₄) that is responsible forincreasing the in vivo serum half-life of the IgG molecule.

In some embodiments, the antibodies of the present invention may beconjugated to a cytotoxic agent. This is any substance that inhibits orprevents the function of cells and/or causes destruction of cells. Theterm is intended to include radioactive isotopes (such as I¹³¹,I¹²⁵,Y⁹⁰, and Re¹⁸⁶), chemotherapeutic agents, and toxins such asenzymatically active toxins of bacterial, fungal, plant, or animalorigin, and fragments thereof. Such cytotoxic agents can be coupled tothe humanized antibodies of the present invention using standardprocedures, and used, for example, to treat a patient indicated fortherapy with the antibody.

A “chemotherapeutic agent” is a chemical compound useful in thetreatment of cancer. There are numerous examples of chemotherapeuticagents that could be conjugated with the therapeutic antibodies of thepresent invention. Examples of such chemotherapeutic agents includealkylating agents such a thiotepa and cyclosphosphamide; alkylsulfonates such as busulfan, improsulfan, and piposulfan; aziridinessuch as benzodopa, carboquone, meturedopa, and uredopa; ethyleniminesand methylamelamines including altretamine, triethylenemelamine,trietylenephosphoramide, triethylenethiophosphoramide, andtrimethylolomelamine; acetogenins (especially bullatacin andbullatacinone); camptothecin (including the synthetic analoguetopotecan); bryostatin; callystatin; CC-1065 (including its adozelesin,carzelesin, and bizelesin synthetic analogues); cryptophycines(particularly cryptophycin 1 and cryptophycin 8); dolastatin,auristatins, (including analogues monomethyl-auristatin E andmonomethyl-auristatin F); duocarmycin (including the syntheticanalogues, KW-2189 and CBI-TMI); eleutherobin; pancratistatin;sarcodictyin; spongistatin; nitrogen mustards such as chlorambucil,chlomaphazine, cholophosphamide, estramustine, ifosfamide,mechlorethamine, mechlorethamine oxide hydrochloride, melphalan,novembichin, phenesterine, prednimustine; trofosfamide, uracil mustard;nitrosureas such as carmustine, chlorozotocin, fotemustine, lomustine,nimustine, ranimustine; antibiotics such as the enediyne antibiotics(e.g., calicheamicin, especially calichemicin gamma1I and calicheamicinphiI1, see for example, Agnew, Chem. Intl. Ed. Engl., 33:183-186;dynemicin, including dynemicin A; bisphosphonates, such as clodronate;esperamicin; as well as neocarzinostatin chromophore and relatedchromoprotein enediyne antibiotic chromomophores), aclacinomysins,actinomycin, authramycin, azaserine, bleomycins, cactinomycin,carabicin, caminomycin, carzinophilin, chromomycins, dactinomycin,daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucine, doxorubicin(Adriamycin™) (including morpholino-doxorubicin,cyanomorpholino-doxorubicin, 2-pyrrolino-doxorubicin, anddeoxydoxorubicin), epirubucin, esorubicin, idarubicin, marcellomycin,mitomycins such as mitomycin C, mycophenolic acid, nogalamycin,olivomycins, peplomycin, potfiromycin, puromycine, quelamycin,rodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex,zinostatin, zorubicin; anti-metabolites such a methotrexate and5-fluorouracil (5-FU); folic acid analogues such as denopterin,methotrexate, pteropterin, trimetrexate; purine analogs such asfludarabine, 6-mercaptopurine, thiamiprine, thioguanine; pyrimidineanalogs such as ancitabine, azacitidine, 6-azauridine, carmofur,cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine;androgens such as calusterone, dromostanolone propionate, epitiostanol,mepitiostane, testolactone; anti-adranals such as aminoglutethimide,mitotane, trilostane; folic acid replenisher such as frolinic acid;aceglatone; aldophosphamide glycoside; aminolevulinic acid; eniluracil;amsacrine; bestrabucil; bisantrene; edatraxate; defofamine; democolcine;diaziquone; elfomithine; elliptinium acetate; an epothilone; etoglucid;gallium nitrate; hydroxyurea; lentinan; lonidamine; maytansinoids suchas maytansine and ansamitocins; mitoguazone, mitoxantrone; mopidamol;nitracrine; pentostatin; phenamet; pirarubicin; losoxantrone;podophyllinic acid; 2-ethylhydrazide; procarbazine; PSK®; razoxane;rhizoxin; sizofuran; spirogermanium; tenuazonic acid; triaziquone;2,2′,2″-trichlorotriethylamine; trichothecenes (especially T-2 toxin,verracurin A, roridin A and anguidine); urethan; vindesine; dacarbazine;mannomustine; mitabronitol; mitolactol; pipobroman; gacytosine;arabinoside (“Ara-C”); cyclophosphamide; thiotepa; taxoids, e.g.,paclitaxel (TAXOL®, Bristol-Myers Squibb Oncology, Princeton, N.J.) anddoxetaxel (TAXOTERE®, Rhone-Poulenc Rorer, Antony, France);chlorambucil; gemcitabine (Gemzar™); 6-thioguanine; mercaptopurine;methotrexate; platinum analogs such as cisplatin and carboplatin;vinblastine; platinum; etoposide (VP-16); ifosfamide; mitoxantrone;vincristine; vinorelbine Navelbine™); novantrone; teniposide;edatrexate; daunomycin; aminopterin; xeloda; ibandronate; CPT-11;topoisomerase inhibitor RFS 2000; difluoromethylornithine (DMFO);retinoids such as retinoic acid; capecitabine; and pharmaceuticallyacceptable salts, acids, or derivatives of any of the above. Alsoincluded in this definition are anti-hormonal agents that act toregulate or inhibit hormone action on tumors such as anti-estrogens andselective estrogen receptor modulators (SERMs), including, for example,tamoxifen (including Nolvadex™) raloxifene, droloxifene,4-hydroxytamoxifen, trioxifene, keoxifene, LY117018, onapristone, andtoremifene (Fareston™); aromatase inhibitors that inhibit the enzymearomatase, which regulates estrogen production in the adrenal glands,such as, for example, 4(5)-imidazoles, aminoglutethimide, megestrolacetate (Megace™), exemestane, formestane, fadrozole, vorozole(Rivisor™), letrozole (Femara™), and anastrozole (Arimidex™); andanti-androgens such as flutamide, nilutamide, bicalutamide, leuprolide,and goserelin; and pharmaceutically acceptable salts, acids, orderivatives of any of the above. Any one or more of these agents may beconjugated to the humanized antibodies of the present invention toprovide a useful therapeutic agent for the treatment of variousdisorders.

The antibodies also may be conjugated to prodrugs. A “prodrug” is aprecursor or derivative form of a pharmaceutically active substance thatis less cytotoxic to tumor cells compared to the parent drug and iscapable of being enzymatically activated or converted into the moreactive form. See, for example, Wilman, 1986, “Prodrugs in CancerChemotherapy”, In Biochemical Society Transactions, 14, pp. 375-382,615th Meeting Belfast and Stella et al., 1985, “Prodrugs: A ChemicalApproach to Targeted Drug Delivery, In: “Directed Drug Delivery,Borchardt et al., (ed.), pp. 247-267, Humana Press. Useful prodrugsinclude, but are not limited to, phosphate-containing prodrugs,thiophosphate-containing prodrugs, sulfate-containing prodrugspeptide-containing prodrugs, D-amino acid-modified prodrugs,glycosylated prodrugs, β-lactam-containing prodrugs, optionallysubstituted phenoxyacetamide-containing prodrugs, and optionallysubstituted phenylacetamide-containing prodrugs, 5-fluorocytosine andother 5-fluorouridine prodrugs that can be converted into the moreactive cytotoxic free drug. Examples of cytotoxic drugs that can bederivatized into a prodrug form include, but are not limited to, thosechemotherapeutic agents described above.

For diagnostic as well as therapeutic monitoring purposes, theantibodies of the invention also may be conjugated to a label, either alabel alone or a label and an additional second agent (prodrug,chemotherapeutic agent and the like). A label, as distinguished from theother second agents refers to an agent that is a detectable compound orcomposition and it may be conjugated directly or indirectly to ahumanized antibody of the present invention. The label may itself bedetectable (e.g., radioisotope labels or fluorescent labels) or, in thecase of an enzymatic label, may catalyze chemical alteration of asubstrate compound or composition that is detectable. Labeled humanizedanti-BAFF antibody can be prepared and used in various applicationsincluding in vitro and in vivo diagnostics.

The antibodies of the present invention may be formulated as part of aliposomal preparation in order to affect delivery thereof in vivo. A“liposome” is a small vesicle composed of various types of lipids,phospholipids, and/or surfactant. Liposomes are useful for delivery to amammal of a compound or formulation, such as a humanized anti-BAFFantibody disclosed herein, optionally, coupled to or in combination withone or more pharmaceutically active agents and/or labels. The componentsof the liposome are commonly arranged in a bilayer formation, similar tothe lipid arrangement of biological membranes.

Certain aspects of the present invention related to isolated nucleicacids that encode one or more domains of the humanized antibodies of thepresent invention. An “isolated” nucleic acid molecule is a nucleic acidmolecule that is identified and separated from at least one contaminantnucleic acid molecule with which it is ordinarily associated in thenatural source of the antibody nucleic acid. An isolated nucleic acidmolecule is distinguished from the nucleic acid molecule as it exists innatural cells.

In various aspects of the present invention one or more domains of thehumanized antibodies will be recombinantly expressed. Such recombinantexpression may employ one or more control sequences, i.e.,polynucleotide sequences necessary for expression of an operably linkedcoding sequence in a particular host organism. The control sequencessuitable for use in prokaryotic cells include, for example, promoter,operator, and ribosome binding site sequences. Eukaryotic controlsequences include, but are not limited to, promoters, polyadenylationsignals, and enhancers. These control sequences can be utilized forexpression and production of humanized anti-BAFF antibody in prokaryoticand eukaryotic host cells.

A nucleic acid sequence is “operably linked” when it is placed into afunctional relationship with another nucleic acid sequence. For example,a nucleic acid presequence or secretory leader is operably linked to anucleic acid encoding a polypeptide if it is expressed as a preproteinthat participates in the secretion of the polypeptide; a promoter orenhancer is operably linked to a coding sequence if it affects thetranscription of the sequence; or a ribosome binding site is operablylinked to a coding sequence if it is positioned so as to facilitatetranslation. Generally, “operably linked” means that the DNA sequencesbeing linked are contiguous, and, in the case of a secretory leader,contiguous and in reading frame. However, enhancers are optionallycontiguous. Linking can be accomplished by ligation at convenientrestriction sites. If such sites do not exist, synthetic oligonucleotideadaptors or linkers can be used.

As used herein, the expressions “cell”, “cell line”, and “cell culture”are used interchangeably and all such designations include the progenythereof. Thus, “transformants” and “transformed cells” include theprimary subject cell and cultures derived therefrom without regard forthe number of transfers.

The term “mammal” for purposes of treatment refers to any animalclassified as a mammal, including humans, domesticated and farm animals,and zoo, sports, or pet animals, such as dogs, horses, cats, cows, andthe like. Preferably, the mammal is human.

A “disorder”, as used herein, is any condition that would benefit fromtreatment with a humanized anti-BAFF antibody described herein. Thisincludes chronic and acute disorders or diseases including thosepathological conditions that predispose the mammal to the disorder inquestion. Non-limiting examples or disorders to be treated hereininclude inflammatory, angiogenic, autoimmune and immunologic disorders,respiratory disorders, cancer, hematological malignancies, benign andmalignant tumors, leukemias and lymphoid malignancies.

The terms “cancer” and “cancerous” refer to or describe thephysiological condition in mammals that is typically characterized byunregulated cell growth. Examples of cancer include, but are not limitedto, carcinoma, lymphoma, blastoma, sarcoma, and leukemia.

As used herein, the term “BAFF-associated disorder” or “BAFF-associateddisease” refers to a condition in which BAFF activity contributes to thedisease and typically where BAFF is abnormally expressed. AnBAFF-associated disorder includes diseases and disorders of the immunesystem, such as autoimmune disorders and inflammatory disorders. Suchconditions include, but are not limited to, rheumatoid arthritis (RA),systemic lupus erythematosus (SLE), scleroderma, Sjogren's syndrome,multiple sclerosis, psoriasis, psoriatic arthritis, inflammatory boweldisease (e.g., ulcerative colitis and Crohn's disease), pulmonaryinflammation, asthma, idiopathic thrombocytopenic purara (ITP) andankylosing spondylitis.

The term “intravenous infusion” refers to introduction of an agent intothe vein of an animal or human patient over a period of time greaterthan approximately 15 minutes, generally between approximately 30 to 90minutes.

The term “intravenous bolus” or “intravenous push” refers to drugadministration into a vein of an animal or human such that the bodyreceives the drug in approximately 15 minutes or less, generally 5minutes or less.

The term “subcutaneous administration” refers to introduction of anagent under the skin of an animal or human patient, preferable within apocket between the skin and underlying tissue, by relatively slow,sustained delivery from a drug receptacle. Pinching or drawing the skinup and away from underlying tissue may create the pocket.

The term “subcutaneous infusion” refers to introduction of a drug underthe skin of an animal or human patient, preferably within a pocketbetween the skin and underlying tissue, by relatively slow, sustaineddelivery from a drug receptacle for a period of time including, but notlimited to, 30 minutes or less, or 90 minutes or less. Optionally, theinfusion may be made by subcutaneous implantation of a drug deliverypump implanted under the skin of the animal or human patient, whereinthe pump delivers a predetermined amount of drug for a predeterminedperiod of time, such as 30 minutes, 90 minutes, or a time periodspanning the length of the treatment regimen.

The term “subcutaneous bolus” refers to drug administration beneath theskin of an animal or human patient, where bolus drug delivery is lessthan approximately 15 minutes; in another aspect, less than 5 minutes,and in still another aspect, less than 60 seconds. In yet even anotheraspect, administration is within a pocket between the skin andunderlying tissue, where the pocket may be created by pinching ordrawing the skin up and away from underlying tissue.

The term “therapeutically effective amount” is used to refer to anamount of an active agent that relieves or ameliorates one or more ofthe symptoms of the disorder being treated. In another aspect, thetherapeutically effective amount refers to a target serum concentrationthat has been shown to be effective in, for example, slowing diseaseprogression. Efficacy can be measured in conventional ways, depending onthe condition to be treated.

The terms “treatment” and “therapy” and the like, as used herein, aremeant to include therapeutic as well as prophylactic, or suppressivemeasures for a disease or disorder leading to any clinically desirableor beneficial effect, including but not limited to alleviation or reliefof one or more symptoms, regression, slowing or cessation of progressionof the disease or disorder. Thus, for example, the term treatmentincludes the administration of an agent prior to or following the onsetof a symptom of a disease or disorder thereby preventing or removing oneor more signs of the disease or disorder. As another example, the termincludes the administration of an agent after clinical manifestation ofthe disease to combat the symptoms of the disease. Further,administration of an agent after onset and after clinical symptoms havedeveloped where administration affects clinical parameters of thedisease or disorder, such as the degree of tissue injury or the amountor extent of metastasis, whether or not the treatment leads toamelioration of the disease, comprises “treatment” or “therapy” as usedherein. Moreover, as long as the compositions of the invention eitheralone or in combination with another therapeutic agent alleviate orameliorate at least one symptom of a disorder being treated as comparedto that symptom in the absence of use of the humanized anti-BAFFantibody composition, the result should be considered an effectivetreatment of the underlying disorder regardless of whether all thesymptoms of the disorder are alleviated or not.

The term “package insert” is used to refer to instructions customarilyincluded in commercial packages of therapeutic products, that containinformation about the indications, usage, administration,contraindications and/or warnings concerning the use of such therapeuticproducts.

Antibodies

In one aspect, anti-BAFF antibodies are described and disclosed. Ofparticular importance for treating autoimmune disease in humans arehumanized anti-BAFF antibodies and compositions disclosed herein. Alsodescribed are binding agents that include an antigen-binding fragment ofan anti-BAFF antibody, in particular a humanized anti-BAFF antibody. Thehumanized anti-BAFF antibodies and binding agents can inhibit theproduction of BAFF-associated cytokines, which contribute to chronicautoimmune and inflammatory diseases. The humanized anti-BAFF antibodiesand binding agents can thus be used in the treatment of a variety ofdiseases or disorders. A humanized anti-BAFF antibody and a BAFF bindingagent each includes at least a portion that specifically recognizes anBAFF epitope (i.e., an antigen-binding fragment).

In the initial characterization of mouse antibodies were selected basedon BAFF receptor binding characterization.

Accordingly in one aspect, an antibody of the present invention has aK_(D) for BAFF, in particular human BAFF, of less than 100 pM. Inanother aspect, an antibody of the present invention has a K_(D) of lessthan 10 pM. In another aspect, an antibody of the present invention hasa K_(D) of less than 1 pM.

The light chain and heavy chain CDRs of the various anti-BAFF antibodiesare shown in Table 3 and Table 4, respectively. Tables 3 and 4 also showfive light chain CDRs and one heavy chain CDR derived from either 1A4 or5B9 mouse antibodies through the humanization process.

TABLE 1 Anti-BAFF Mouse Leads - Vκ Sequences Designation Sequence206G9A10 GACATTGTGATGACCCAGTCTCAAAAATTCATGTCCACAACAGTAGGAGACAGGGTCAGCATCACCTGCAAGGCCAGTCAGAATGCGGGTGCTGCTGTAGCCTGGTTTCAACAGAAACCAGGACAATCTCCTAAACTACTGATTTACTCAGCATCCAATCGGTATACTGGAGTCCCTGATCGCTTCACAGGCAGTGGATCGGGGACAGATTTCACTCTCACCATTAGCAATGTGCAGTCTGAGGACCTGGCAGATTATATCTGTCAACAATACAGAAGCTATCCTCGGACGTTCGGAGGAGGCACCAAGCTGGAAATCAAA (SEQ ID NO: 40)DIVMTQSQKFMSTTVGDRVSITCKASQNAGAAVAWFQQKPGQSPKLLIYSASNRYTGVPDRFTGSGSGTDFTLTISNVQSEDLADYICQQYRSYPRTFGGGTKLEIK (SEQ ID NO: 41) 227D5A7GACATTGTGATGACCCAGTCTCAAAAATTCATGTCCACAACAGTAGGAGACAGGGTCAGCATCACCTGCAAGGCCAGTCAGAATGCGGGTGCTGCTGTAGCCTGGTTTCAACAGAAACCGGGACAATCTCCTAAATTACTGATTTACTCAGCATCCAATCGGTATACTGGAGTCCCTGATCGCTTCACAGGCAGTGGATCGGGGACAGATTTCACTCTCACCATTAGCAATGTGCAGTCTGAGGACCTGGCAGATTATATCTGTCAACAATACAGAAGCTTTCCTCGGACGTTCGGAGGAGGCACCAAGCTGGAAATCAAA (SEQ ID NO: 42)DIVMTQSQKFMSTTVGDRVSITCKASQNAGAAVAWFQQKPGQSPKLLIYSASNRYTGVPDRFTGSGSGTDFTLTISNVQSEDLADYICQQYRSFPRTFGGGTKLEIK (SEQ ID NO: 43) 250E5A11GACATTGTGATGACCCAGTCTCAAAAATTCATGTCCACAACAGTAGGAGACAGGGTCAGCATCACCTGCAAGGCCAGTCAGAATGCGGGTGCTGCTGTAGCCTGGTTTCAACAGAAACCAGGACAATCTCCTAAACTACTGATTTACTCAGCATCCAATCGGTATACTGGAGTCCCTGATCGCTTCACAGGCAGTGGATCGGGGACAGATTTCACTCTCACCATTACCAATGTGCAGTCTGAGGACCTGGCAGATTATATCTGTCAACAATACAGAAGCTTTCCTCGGACGTTCGGAGGAGGCACTAAGCTGGAAATCAAA (SEQ ID NO: 44)DIVMTQSQKFMSTTVGDRVSITCKASQNAGAAVAWFQQKPGQSPKLLIYSASNRYTGVPDRFTGSGSGTDFTLTITNVQSEDLADYICQQYRSFPRTFGGGTKLEIK (SEQ ID NO: 45) 227D3B11GACATTGTGATGACCCAGTCTCAAAAAATCATGTCCACAACAGTGGGAGACAGGGTCAGCATCACCTGCAAGGCCAGTCAGAATGCGGGTATTGATGTAGCCTGGTTTCAACAGAAACCAAGACAATCTCCTAAACTACTGATTTTCTCAACATCCAATCGATATACTGGAGTCCCAGATCGCTTCGCAGGCAGTGGATCGGGGACAGATTTCACTCTCACCATTTACAATGTGCAGTCTGAAGACCTGGCAGATTATTTCTGTCTGCAATATAGAAGTTATCCTCGGACGTTCGGAGGGGGGACCAAGCTGGAAATAAAA (SEQ ID NO: 46)DIVMTQSQKIMSTTVGDRVSITCKASQNAGIDVAWFQQKPRQSPKLLIFSTSNRYTGVPDRFAGSGSGTDFTLTIYNVQSEDLADYFCLQYRSYPRTFGGGTK LEIK (SEQ ID NO: 47)235F5B9 GACATTGTGATGACCCAGTCTCAAAAAATCATGTCCACAACAGTGGGAGACAGGGTCAGCATCACCTGCAAGGCCAGTCAGAATGCGGGTATTGATGTAGCCTGGTTTCAACAGAAACCAAGACAATCTCCTAAACTACTGATTTTCTCAAAATCCAATCGATATACTGGAGTCCCAGATCGCTTCGCAGGCAGTGGATCGGGGACAGATTTCACTCTCACCATTTACAATGTGCAGTCTGAAGACCTGGCAGATTATTTCTGTCTGCAATATAGAAGTTATCCTCGGACGTTCGGAGGAGGCACCAAGCTGGAAATCAAA (SEQ ID NO: 48)DIVMTQSQKIMSTTVGDRVSITCKASQNAGIDVAWFQQKPRQSPKLLIFSKSNRYTGVPDRFAGSGSGTDFTLTIYNVQSEDLADYFCLQYRSYPRTFGGGTK LEIK (SEQ ID NO: 49)217H12A7 GACATTGTGATGACCCAGTCTCAAAAATTCATGTCCACAACAGTAGGAGACAGGGTCAGCATCACCTGCAAGGCCAGTCAGAATGCGGGTACTGCTGTAGCCTGGTTTCAACAGAAACCAGGACAATCTCCTAAACTACTGATTTACTCAGCATTTAATCGGTATACTGGAGTCCCTGATCGCTTCACAGGCAGTGGATCGGGGACAGATTTCACTCTCACCATTAGCAATATGCAGTCTGAAGACCTGGCAGATTATATCTGTCAACAATATAGAAGCTATCCTCGGACGTTCGGAGGAGGCACCAAGCTGGAAATCAAA (SEQ ID NO: 50)DIVMTQSQKFMSTTVGDRVSITCKASQNAGTAVAWFQQKPGQSPKLLIYSAFNRYTGVPDRFTGSGSGTDFTLTISNMQSEDLADYICQQYRSYPRTFGGGTKLEIK (SEQ ID NO: 51) 210D9B8GACATTGTGATGACCCAGTCTCAAAAATTCGTGTCCACAACACTAGGGGACAGGGTCAGCATCACCTGCAAGGCCAGTCAGAGTGTGGGTATTGCTGTAGCCTGGTATCAACAGAAACCAGGACATTCTCCTAACCTACTGATTTTCTCAACATCCAATCGCTACACTGGAGTCCCTGATCGCTTCACAGGCAGCGGATCTGGGACAGATTTCACTCTCACCATTAGCGATGTGCAGTCTGAAGACCTGGCAGATTATTTCTGTCAGCAATATAGCAGGTATCCTCGGACGTTCGGTGGAGGCACCAAGCTGGAGATCAAA (SEQ ID NO: 52)DIVMTQSQKFVSTTLGDRVSITCKASQSVGIAVAWYQQKPGHSPNLLIFSTSNRYTGVPDRFTGSGSGTDFTLTISDVQSEDLADYFCQQYSRYPRTFGGGTK LEIK (SEQ ID NO: 53)214G4B7 GACATTGTGATGACCCAGTCTCAAAAATTCATGTCCACAACAGTAGGAGACAGGGTCAGCATCACCTGCAAGGCCAGTCAGAATGCGGGTACTGCTGTAGCCTGGTTTCAACAGAAACCAGGACAATCTCCTAAACTACTGATTTTCTCAACATCCAATCGGTATACTGGAGTCCCTGATCGCTTCACAGGCAGTGGATCGGGGACAGATTTCACTCTCACCATTAGCAATATGCAGTCTGAAGACCTGGCAGATTATTTCTGTCTGCAATATAGAAGCTATCCTCGGACGTTCGGAGGAGGCACCAAGCTGGAAATCAAA (SEQ ID NO: 54)DIVMTQSQKFMSTTVGDRVSITCKASQNAGTAVAWFQQKPGQSPKLLIFSTSNRYTGVPDRFTGSGSGTDFTLTISNMQSEDLADYFCLQYRSYPRTFGGGTKLEIK (SEQ ID NO: 55) 13J018-1A4GACATCCAGATGACCCAGTCTCCATCCTCATTATCTGCCTCTCTGGGAGAAAGAGTCAGTCTCACTTGTCGGGCAAGTCAAGACATTGGTAATAGGTTAAACTGGCTTCAGCAGGAACCAGATGGAACTATTAAACGCCTGATCTACGCCACATCCAGTTTAGATTCTGGTGTCCCCAAAAGGTTCAGTGGCAGTAGGTCTGGGTCGGATTATTCTCTCACCATCAGCAGCCTTGAGTCTGAAGATTTTGTAGACTATTACTGTCTACAATATGCTAGTTCTCCATTCACGTTCGGCACGGGGACAAAATTGGAAATAAAA (SEQ ID NO: 56)DIQMTQSPSSLSASLGERVSLTCRASQDIGNRLNWLQQEPDGTIKRLIYATSSLDSGVPKRFSGSRSGSDYSLTISSLESEDFVDYYCLQYASSPFTFGTGTKLEI K (SEQ ID NO: 57)1002E8A6 GACATCAAAATGACCCAGTCTCCATCTTCCATGTATGCATCTCTAGGAGAGAGAGTCACTATCACTTGCAAGGCGAGTCAGGACATTAATAGCTATTTAACCTGGTTCCAGCAGAAACCAGGGAAATCTCCTGAGACCCTGATCTATCGTGCAAACAGATTGGTATCTGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGCAAGATTATTCTCTCACCATCAGCAGCCTGGAATATGAAGATATGGGAATTTATTCTTGTCTACAGTATGATGAGTTTCCGTACACGTTCGGAGGGGGGACCAAGCTGGAAATAAAA (SEQ ID NO: 118)DIKMTQSPSSMYASLGERVTITCKASQDINSYLTWFQQKPGKSPETLIYRANRLVSGVPSRFSGSGSGQDYSLTISSLEYEDMGIYSCLQYDEFPYTFGGGTKLEIK (SEQ ID NO: 119) 1070A6B7GATGTTGTGATGACCCAAACTCCACTCTCCCTGCCTGTCAGTCTTGGAGATCAAGCCTCCATCTCTTGCAGATGTAGTCAGAGCCTTGTACACAGTAATGGAAACACGTATTTACATTGGTACCTGCAGAAGCCAGGCCAGTCTCCAAAGCTCCTGATCTACAAAGTTTCCGACCGATTTTCTGGGGTCCCAGACAGGTTCAGTGGCAGTGGATCAGGGACAGATTTCACACTCAGGATCAGCAGAGTGGAGGCTGACGATCTGGGAGTTTATTTCTGCTCTCAAAGTACACATGTTCCGCTCACGTTCGGTGCTGGGACCAAGCTGGAGCTGAAA (SEQ ID NO: 120)DVVMTQTPLSLPVSLGDQASISCRCSQSLVHSNGNTYLHWYLQKPGQSPKLLIYKVSDRFSGVPDRFSGSGSGTDFTLRISRVEADDLGVYFCSQSTHVPLTFGAGTKLELK (SEQ ID NO: 121) 1094C4E6GACATTGTGATGACCCAGTCTCACAAATTCATGTCCACATCAGTAGGAGACAGGGTCACCATCACCTGCAAGGCCAGTCAGGATGTGGCTACTGCTGTAGCCTGGTATCAACAGAAACCAGGGCAATCTCCTAAACTACTAATTTACTGGGCATCCACCCGGCACACTGGAGTCCCTGATCGCTTCACAGGCAGTGGATCTGGGACAGATTTCACTCTCACCATTAGCAATGTGCAGTCTGAAGACTTGGCAAATTATTTCTGTCAGCAATATAGCAACTATCCGTACACGTTCGGAGGGGGGACCACGCTGGAAATAAAA (SEQ ID NO: 122)DIVMTQSHKFMSTSVGDRVTITCKASQDVATAVAWYQQKPGQSPKLLIYWASTRHTGVPDRFTGSGSGTDFTLTISNVQSEDLANYFCQQYSNYPYTFGGGTTLEIK (SEQ ID NO: 123) 27I21-3C7GACATCCAGATGACCCAGTCTCCATCCTCCTTATCTGCCTCTCTGGGAGAAAGAGTCAGTCTCACTTGTCGGGCAAGTCAGGACATTGGTAATAGGTTAAACTGGCTTCAGCAGGCACCAGATGGAACTATTAAACGCCTGATCTACGCCACATCCAGTTTAGATTCTGGTGTCCCCAAAAGGTTCAGTGGCAGTCGGTCTGGGTCAGATTATTCTCTCACCATCAGCAGCCTTGAATCTGAAGATTTTGTAGACTATTACTGTCTACAATATGCTAGTTATCCATTCACGTTCGGCACGGGGACAAAATTGGAAATAAAA (SEQ ID NO: 124)DIQMTQSPSSLSASLGERVSLTCRASQDIGNRLNWLQQAPDGTIKRLIYATSSLDSGVPKRFSGSRSGSDYSLTISSLESEDFVDYYCLQYASYPFTFGTGTKLEIK (SEQ ID NO: 125) 317H2A6GACATTGTGATGACCCAGTCTCAAAAATTTTTGTCCACAACAATAGGAGACAGGGTCAGCATCACCTGCAAGGCCAGTCAGAATGTGGGTTCTGCTGTAGTCTGGTATCAACAGAAACCAGGCCAACCTCCTAAACTACTGATTACCTCAGCATCCAATCGGTACAGTGGAGTCCCAGATCGCTTCACAGGCAGTGGATCTGGGACAGATTTCACTCTCACCGTTAGCAATGTGCAGTCTGTAGACCTGGCAGATTATTTCTGTCAACAATATAGCAACTATCCTCTCACGTTCGGTGCTGGGACCAAGCTGGAGCTGAAA (SEQ ID NO: 126)DIVMTQSQKFLSTTIGDRVSITCKASQNVGSAVVWYQQKPGQPPKLLITSASNRYSGVPDRFTGSGSGTDFTLTVSNVQSVDLADYFCQQYSNYPLTFGAGTKLELK (SEQ ID NO: 127) 319B8A12GACATTGTGATGACCCAGTCTCAAAAATTTGTGTCGACAAGAGTTGGAGACAGGGTCAGCATCACCTGCAAGGCCAGTCAGAATGTGGGCGCTGCTGTAGTCTGGTATCAACAGAAATCAGGCCAACCTCCTAAACTACTGATTAGGTCAGCATCCAATCGGTACATTGGAGTCCCTGATCGCTTCACAGGCAGTGGGTCTGGGACAGATTTCACTCTCACCGTTAGCGATGTGCAGTCTGGAGACCTGGCAGATTATTTCTGTCAGCAATATAGCAACTATCCTCTCACGTTCGGTGCTGGGACCAAGCTGGAACTGACACGGGCTGAT (SEQ ID NO: 128)DIVMTQSQKFVSTRVGDRVSITCKASQNVGAAVVWYQQKSGQPPKLLIRSASNRYIGVPDRFTGSGSGTDFTLTVSDVQSGDLADYFCQQYSNYPLTFGAGTKLELTRAD (SEQ ID NO: 129) 320F9C5GACATTGTGATGACCCAGTCTCAAAAATTCATGTCCACAACAGTAGGAGACAGGGTCAGCATCACCTGCAAGGCCAGTCAGAATGTGGGTAGTGTTGTAGCCTGGTATCAACAGAGACCAGGACAATCTCCTACACTACTGATTTACTCAGCATCCAATCGGTACACTGGAGTCCCTGATCGCTTCACTGGCAGTGGATCTGGGACAGATTTCACTCTCACCATTAGCAATATGCAGTCGGAAGACCTGGCAGATTATTTCTGTCAGCAATATAGCAGCTATCCTCTCACGTTCGGTGCTGGGACCAAGCTGGAGCTGAAA (SEQ ID NO: 130)DIVMTQSQKFMSTTVGDRVSITCKASQNVGSVVAWYQQRPGQSPTLLIYSASNRYTGVPDRFTGSGSGTDFTLTISNMQSEDLADYFCQQYSSYPLTFGAGTKLELK (SEQ ID NO: 131) 323E9D1GACATTGTGATGACCCAGTCTCAAAAATTTGTGTCGACAAGAGTTGGAGACAGGGTCAGCATCACCTGCAAGGCCAGTCAGAATGTGGGCGCTGCTGTAGTCTGGTATCAACAGAAATCAGGCCAACCTCCTAAACTACTGATTAGGTCAGCATCCAATCGGTACATTGGAGTCCCTGATCGCTTCACAGGCAGTGGGTCTGGGACAGATTTCACTCTCACCGTTAGCGATGTGCAGTCTGGAGACCTGGCAGATTATTTCTGTCAGCAATATAGTAACTATCCTCTCACGTTCGGTGCTGGGACCAAGCTGGAACTGACA (SEQ ID NO: 132)DIVMTQSQKFVSTRVGDRVSITCKASQNVGAAVVWYQQKSGQPPKLLIRSASNRYIGVPDRFTGSGSGTDFTLTVSDVQSGDLADYFCQQYSNYPLTFGAGTKLELT (SEQ ID NO: 133) 332C1B12GACATTGTGCTGACACAGTCTCCTGCTTCCTTACCTGTTTCTCTGGGGCAGAGGGCCACCATCTCCTGCAGGGCCAGCAAAGGTGTCAGTACATCTAGCTATACTTTCATTCACTGGTACCAACAGAAACCTGGACAGCCGCCCAAACTCCTCATCAAGTATGCATCCAACCTAGAATCTGGGGTCCCTGCCAGGTTCAGTGGCAGTGGGTCTGGGACAGACTTCACCCTCAACATCCATCCTGTGGAGGAGGAGGATGTTGCAACATATTACTGTCAGCACAGTAGGGAGTTTCCTCGGACGTTCGGTGGAGGCACCAAGCTGGAAATCAAA (SEQ ID NO: 134)DIVLTQSPASLPVSLGQRATISCRASKGVSTSSYTFIHWYQQKPGQPPKLLIKYASNLESGVPARFSGSGSGTDFTLNIHPVEEEDVATYYCQHSREFPRTFGGGTKLEIK (SEQ ID NO: 135) 344B9D9GACATCCAGATGACTCAGTCTCCAGCCTCCCTATCTGCATCTGTGGGAGAAACTGTCACCATCACATGTCGAGCAAGTGGGAATATTCACAATTATTTAGCATGGTATCAGCAGAAACAGGGAAAATCTCCTCAGCTCCTGGTCTATAGTGCAATAACCTTAGCAGATGGTGTGCCATCAAGGTTCAGTGGCAGTGGATCAGAAACACAATTTTCTCTCAAGATCAACAGCCTGCAGCCTGAAGATTTTGGGATTTATTACTGTCAACATTTTTGGAATACTCCGTACACGTTCGGAGGGGGGACCAAGCTGGAAATAAAA (SEQ ID NO: 136)DIQMTQSPASLSASVGETVTITCRASGNIHNYLAWYQQKQGKSPQLLVYSAITLADGVPSRFSGSGSETQFSLKINSLQPEDFGIYYCQHFWNTPYTFGGGTKLEIK (SEQ ID NO: 137) 348A6C1GACATTGTGATGACCCAGTCTCAAAAATTTATGTCCACAACAGTTGGAGACAGGGTCAGCATCACCTGCAAGGCCAGTCAGAATGTGGGTGCTGCTGTAGCCTGGTATCAACAGAAACCAGGCCAACCTCCTAAACTACTGATTAGGTCAGCATCCAATCGGTACATTGGAGTCCCTGATCGCTTCACAGGCAGTGGGTCTGGGACAGATTTCACTCTCACCGTTAGCGATGTGCAGTCTGTAGACCTGGCAGATTATTTCTGTCAGCAATATAGCAACTATCCTCTCACGTTCGGTGCTGGGACCAAGCTGGAACTGACACGGGCTGAT (SEQ ID NO: 138)DIVMTQSQKFMSTTVGDRVSITCKASQNVGAAVAWYQQKPGQPPKLLIRSASNRYIGVPDRFTGSGSGTDFTLTVSDVQSVDLADYFCQQYSNYPLTFGAGTKLELTRAD (SEQ ID NO: 139) 352G11A10GACATCAAGATGACCCAGTCTCCATCTTCCATATATGCATCTCTAGGAGAGAGAGTCACTATCACTTGCAAGGCGAGTCAGGACATTCATAGCTATTTAAGTTGGTTCCAGCAGAAACCAGGGAAATCTCCTAAGACCCTGATGTATCGTACAAATAGATTGGTAGATGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGCAAGATTATTCTCTCACCATCAGGAGCCTGGAATATGAAGATATGGGAAATTATTATTGTCTACAGTATGATGAATTTCCGTACACGTTCGGCGGGGGGGCCAAGTTGGAAGTAAAA (SEQ ID NO: 140)DIKMTQSPSSIYASLGERVTITCKASQDIHSYLSWFQQKPGKSPKTLMYRTNRLVDGVPSRFSGSGSGQDYSLTIRSLEYEDMGNYYCLQYDEFPYTFGGGAKLEVK (SEQ ID NO: 141) 363D4A10GACATTGTGATGACCCAGTCTCAAAAATTCATGTCCACAACAGTAGGAGACAGGGTCACCATCACCTGCAAGGCCAGTCAGAATGTGGGTAGTGCTGTAGTCTGGTATCAACAGAAACCAGGACAATCTCCTATATTACTGATTTTCTCAGCATCCAATCGGTACACTGGAGTCCCTGATCGCATCACAGGCAGTGGGTCTGGGGCAGAATTCACTCTCACCATTAGCAGTGTGCAGTCTGAAGACCTGGCAGAATATTTCTGTCAGCAATATAGCAGCTATCCTCTCACGTTCGGTGCTGGGACCAAGCTGGAGCTGAAA (SEQ ID NO: 142)DIVMTQSQKFMSTTVGDRVTITCKASQNVGSAVVWYQQKPGQSPILLIFSASNRYTGVPDRITGSGSGAEFTLTISSVQSEDLAEYFCQQYSSYPLTFGAGTKLELK (SEQ ID NO: 143) 381A6A9GACATCAAGATGACCCAGTCTCCATCTTCCATATATGCATCTCTAGGAGAGAGAGTCACTATCACTTGCAAGGCGAGTCAGGACATTAATAGCTATTTAAGCTGGTTCCAGCAGAAACCAGGGAAATCTCCTAAGACCCTGATGTATCGTGCAAACAGATTGGTAGATGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGCAAGATTATTCTCTCACCATCAGCAGCCTGGAATATGAAGATATGGGAAATTATTATTGTCTACAGTATGATGAGTTTCCGTACACGTTCGGAGGGGGGGCCAAGCTGGAAATAAAA (SEQ ID NO: 144)DIKMTQSPSSIYASLGERVTITCKASQDINSYLSWFQQKPGKSPKTLMYRANRLVDGVPSRFSGSGSGQDYSLTISSLEYEDMGNYYCLQYDEFPYTFGGGAKLEIK (SEQ ID NO: 145) 384D5A2GACATCCAGATGACTCAGTCTCCAGCCTCCCTATCTGTATCTGTGGGAGAAACTGTCACCATCACATGTCGATCAAGTGAGAATATTTACAGTAGTTTAGCATGGTATCAACAGAAACAGGGAAAATCTCCTCAGCTCCTGGTCTATGCTGCAACAAACTTAGCAAAAGGTGTGCCGTCAAGGTTCAGTGGCAGTGGATCAGGCACACAGTATTCCCTCAAGATCAACAGCCTACAGTCTGAAGATTTTGGGAGTTATTTCTGTCAACATTTTTGGGGTAGTCCATTCGCGTTCGGCTCGGGGACAAAGTTGGAAATAAAA (SEQ ID NO: 146)DIQMTQSPASLSVSVGETVTITCRSSENIYSSLAWYQQKQGKSPQLLVYAATNLAKGVPSRFSGSGSGTQYSLKINSLQSEDFGSYFCQHFWGSPFAFGSGTKLEIK (SEQ ID NO: 147) 394F5A5GACATTGTGATGACCCAGTCTCAAAAATTTATGTCCACAACAATAGGAGACAGGGTCAGCATCACCTGCAAGGCCAGTCAGAATGTGGGTTCTGCTGTGGCCTGGTATCAACAGAAACCAGGACAACCTCCCAAACTACTGATTTACTCAACATCCAATCGGTACACTGGAGTCCCTGATCGCTTCACAGGCAGTAGATCTGGGACAGATTTCACTCTCACCGTTAGCAATATGCAGTCTGAAGACCTGGCAGATTATTTCTGTCAGCAATATGCCAGCTATCCTCTCACATTCGGTACTGGGACCAAGCTGGAGCTGAAA (SEQ ID NO: 148)DIVMTQSQKFMSTTIGDRVSITCKASQNVGSAVAWYQQKPGQPPKLLIYSTSNRYTGVPDRFTGSRSGTDFTLTVSNMQSEDLADYFCQQYASYPLTFGTGTKLELK (SEQ ID NO: 149) 409F12A11GACATTGTGCTGACACAGTCTCCTGCTTCCTTAGCTTTATCTCTGGGGCAGAGGGCCACCATCTCATGCAGGGCCACCAAAGGGGTCAGTAAATCTGGCTATAGTTATATGCACTGGTACCAACAGAAACCAGGGCAGCCACCCAAACTCCTCATCTATCTTGCATCCAACCTAGAATCTGGGGTCCCTGCCAGGTTCAGTGGCAGTGGGTCTGGGACAGACTTCACCCTCAATATCCATCCTGTGGAGGAGGAGGATGTTGCAACCTATTACTGTCAGCACAGTAGGGAGCTTCCGCTCACGTTCGGTGCTGGGACCAAGCTGGAGCTGAAA (SEQ ID NO: 150)DIVLTQSPASLALSLGQRATISCRATKGVSKSGYSYMHWYQQKPGQPPKLLIYLASNLESGVPARFSGSGSGTDFTLNIHPVEEEDVATYYCQHSRELPLTFGAGTKLELK (SEQ ID NO: 151) 418F6D9ATTGTGCTGACCCAATCTTCAGCTTCTTTGGCTGTGTCTCTAGGGCAGAGGGCCACCATATCCTGCAGAGCCAGTGAAAGTGTTGATAGTTATGGCAATAGTCTTATGCACTGGTACCAGCAGAAACCAGGACAGCCACCCAAACTCCTCATCTATATTGCATCCAACCTAGAATCTGGGGTCCCTGCCAGGTTCAGTGGCAGTGGGTCTAGGACAGACTTCACCCTCACCATTGATCCTGTGGAGGCTGATGATGCTGCAACCTATTACTGTCAGCAAAATAGTGAGGATCCTCGGACGTTCGGTGGAGGCACCAAGCTGGAAATCAAA (SEQ ID NO: 152)IVLTQSSASLAVSLGQRATISCRASESVDSYGNSLMHWYQQKPGQPPKLLIYIASNLESGVPARFSGSGSRTDFTLTIDPVEADDAATYYCQQNSEDPRTFGGGTKLEIK (SEQ ID NO: 153) 431G5A3AAAATTGTGCTGACCCAATCTTCAGCTTCTTTGGCTGTGTCTCTAGGGCAGAGGGCCACCATATCCTGCAGAGCCAGTGAAAGTGTTGATCGTTATGGCAATAGTCTTATGCACTGGTACCAGCAGAAACCAGGACAGCCACCCAAACTCCTCATCTATATTGCATCCAACCTAGAATCTGGGGTCCCTGCCAGGTTCAGTGGCAGTGGGTCTAGGACAGACTTCACCCTCACCATTGATCCTGTGGAGGCTGATGATGCTGCAACCTATTACTGTCAGCAAAATAATGAGGATCCTCGGACGTTCGGTGGAGGCACCAAGCTGGAAATCAAA (SEQ ID NO: 154)KIVLTQSSASLAVSLGQRATISCRASESVDRYGNSLMHWYQQKPGQPPKLLIYIASNLESGVPARFSGSGSRTDFTLTIDPVEADDAATYYCQQNNEDPRTFGGGTKLEIK (SEQ ID NO: 155) 435A6B3GACATCAAGATGACCCCGTCTCCTTCTTCCATGTATGCATCTCTCGGAGAGAGAGTCACTATCACTTGCAAGGCGAGTCAGGACATTAATAGATATTTAAGCTGGTTCCAGCAGAAACCAGGGAAATCTCCTAAGACCCTGATCTATCGTGCAAATAGATTGGTAGATGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGGCAAGATTACTCTCTCACCATCAGCAGCCTGGAGTATGAAGATATGGGAATTTATTATTGTCTACAGTATGATGAATTTCCTTACACGTTCGGAGGGGGGACCAAGCTGGAAATAAAA (SEQ ID NO: 156)DIKMTPSPSSMYASLGERVTITCKASQDINRYLSWFQQKPGKSPKTLIYRANRLVDGVPSRFSGSGSGQDYSLTISSLEYEDMGIYYCLQYDEFPYTFGGGTKLEIK (SEQ ID NO: 157) 436H2C12AACATTGTGCTGACCCAATCTCCAGCTTCTTTGGCTGTGTCTCTAGGGCAGAGGGCCACCATATCCTGCAGAGCCAGTGAAAGTGTTGATAATTATGGCAATAGTTTTATGCACTGGTACCAGCAGAAACCAGGACAGCCACCCAAACTCCTCATCTCTCTTGCATCCAACCTAGAATCTGGGGTCCCTGCCAGGTTCAGTGGCAGTGGGTCTAGGACAGACTTCACCCTCACCATTGATCCTGTGGAGGCTGATGATGCTGCAACCTATTACTGTCAGCAAAATAATGAGGATCCTCGGACGTTCGGTGGAGGCACCAAGCTGGAAATCAAA (SEQ ID NO: 158)NIVLTQSPASLAVSLGQRATISCRASESVDNYGNSFMHWYQQKPGQPPKLLISLASNLESGVPARFSGSGSRTDFTLTIDPVEADDAATYYCQQNNEDPRTFGGGTKLEIK (SEQ ID NO: 159) 436H6A9GACATTGTGCTGACACAGTCTCCTGCTTCCTTAGGTGTATCTCTGGGGCAGAGGGCCACCATCTCTTGCAGGGCCACCAAAGGGGTCACTAAATCTGGCTATAGTTATATTCACTGGTACCAACAGAAACCAGGACAGCCACCCAAACTCCTCATCTATCTTGCATCCAACCTACAATCTGGGGTCCCTGCCAGGTTCAGTGGCAGTGGGTCTGGGACAGACTTCACCCTCAACATCCATCCGGTGGAGGAGGAGGATGCTGCAACCTATTACTGTCAGCACAGTAGGGAGCTTCCGCTCACGTTCGGTGCTGGGACCAAGCTGGAGCTGAAA (SEQ ID NO: 160)DIVLTQSPASLGVSLGQRATISCRATKGVTKSGYSYIHWYQQKPGQPPKLLIYLASNLQSGVPARFSGSGSGTDFTLNIHPVEEEDAATYYCQHSRELPLTFGAGTKLELK (SEQ ID NO: 161) 440E9D12AACATTGTGCTGACCCAATCTCCAGCTTCTTTGCCTGTGTCTCTAGGGCAGAGGGCCACCATGTCCTGCAGAGCCAGTAAAAGTGTTGATAGTTATGGCACTAGTTTTATGCACTGGTACCAACACAGACCAGGACAGCCACCCAAACTCCTCATCTCTCTTGCATCCAACCTAGAATCTGGGGTCCCTGGCAGGTTCAGTGGCAGTGGGTCTAGGACAGACTTCACCCTCACCATTGATCCTGTGGAGCCTGATGATGCTGCAACCTATTACTGTCAACAAAATAATGAGGATCCTCGGACGTTCGGTGGAGGCACCACGCTGGAAATCAAA (SEQ ID NO: 162)NIVLTQSPASLPVSLGQRATMSCRASKSVDSYGTSFMHWYQHRPGQPPKLLISLASNLESGVPGRFSGSGSRTDFTLTIDPVEPDDAATYYCQQNNEDPRTFGGGTTLEIK (SEQ ID NO: 163) 441E6F2AACATTGTGTTGACCCAATCTCCAGCTTCTTTGGCTGTGTCTCTAGGACAGAGGGCCACCATATCCTGCAGAACCAGTGAAAGTGTTGATAGTTATGGCAATAGTTTTATGTTCTGGTTCCAGCAGAAACCAGGACAGGCACCCAAACTCCTCATCTTTCTTACATCCAACCTCGAATCTGGGGTCCCTGCCAGGTTCAGTGGCAGTGGGTCTAGGACAGACTTCACCCTCACCATTGATCCTGTGGAGGCTGATGATGCTGCAACCTATTACTGTCAGCAAAGTAATGAGGATCCTCGGACGTTCGGTGGAGGCACCAAGCTGGAAATCAAA (SEQ ID NO: 164)NIVLTQSPASLAVSLGQRATISCRTSESVDSYGNSFMFWFQQKPGQAPKLLIFLTSNLESGVPARFSGSGSRTDFTLTIDPVEADDAATYYCQQSNEDPRTFGGGTKLEIK (SEQ ID NO: 165) 443C11A12GACATCAAGATGACCCCGTCTCCTTCTTCCATGTATGCATCTCTCGGAGAGAGAGTCACTATCACTTGCAAGGCGAGTCAGGACATTAATAGCTATTTAAGTTGGTTCCAGCAGAAACCAGGGAAATCTCCTAAGACCCTGATCTATCGTGCAAATAGATTGGTAGATGGGGTCCCATCAAGGTTCAGTGGCAGTGGATCTGGCCAAGATTACTCTCTCACCATCAGCAGCCTGGAATATGAAGATATGGGAATTTATTATTGTCTACAGTATGATGAATTTCCTTACACGTCCGGAGGGGGGACCAAGCTGGAAATAAAG (SEQ ID NO: 166)DIKMTPSPSSMYASLGERVTITCKASQDINSYLSWFQQKPGKSPKTLIYRANRLVDGVPSRFSGSGSGQDYSLTISSLEYEDMGIYYCLQYDEFPYTSGGGTKLEIK (SEQ ID NO: 167) 444G1A10AACATTGTGTTGACCCAATCTCCAGCTTCTTTGGCTGTGTCTCTAGGACAGAGGGCCACCATATCCTGCAGAGCCAGTGAAAGTGTTGATAGTTATGGCAATAGTTTTATGTTCTGGTTCCAGCAGAAACCAGGACAGGCACCCAAACTCCTCATCTTTCTTACATCCAACCTCGAATCTGGGGTCCCTGCCAGGTTCAGTGGCAGTGGGTCTAGGACAGACTTCACCCTCACCATTGATCGTGTGGAGGCTGATGATGCTGCAACCTATTACTGTCAGCAAAGTAATGAGGATCCTCGGACGTTCGGTGGAGGCACCAAGCTGGAAATCAAA (SEQ ID NO: 168)NIVLTQSPASLAVSLGQRATISCRASESVDSYGNSFMFWFQQKPGQAPKLLIFLTSNLESGVPARFSGSGSRTDFTLTIDRVEADDAATYYCQQSNEDPRTFGGGTKLEIK (SEQ ID NO: 169) 450A2A7ATTGTGCTGACCCAATCTTCAGCTTCTTTGGCTGTGTCTCTAGGGCAGAGGGCCACCATATCCTGCAGAGCCAGTGAAAGTGTTGATCGTTATGGCAATAGTCTTATGCACTGGTACCAGCAGAAACCAGGACAGCCACCCAAACTCCTCATCTATATTGCATCCAACCTAGAATCTGGGGTCCCTGCCAGGTTCAGTGGCAGTGGGTCTAGGACAGACTTCACCCTCACCATTGATCCTGTGGAGGCTGATGATGCTGCAACCTATTACTGTCAGCAAAATAATGAGGATCCTCGGACGTTCGGTGGAGGCACCAAGCTGGAAATCAAA (SEQ ID NO: 170)IVLTQSSASLAVSLGQRATISCRASESVDRYGNSLMHWYQQKPGQPPKLLIYIASNLESGVPARFSGSGSRTDFTLTIDPVEADDAATYYCQQNNEDPRTFGGGTKLEIK (SEQ ID NO: 171) 456H11B7AACATTGTGCTGACCCAATCTCCAGCTTCTTTGCCTGTGTCTCTAGGGCAGAGGGCCACCATGTCCTGCAGAGCCAGTAAAAGTGTTGATAGTTATGGCACTAGTTTTATGCACTGGTACCAACACAGACCAGGACAGCCACCCAAACTCCTCATCTCTCTTGCATCCAACCTAGAATCTGGGGTCCCTGGCAGGTTCAGTGGCAGTGGGTCTAGGACAGACTTCACCCTCACCATTGATCCTGTGGAGCCTGATGATGCTGCAACCTATTACTGTCAACAAAATAATGAGGATCCTCGGACGTTCGGTGGAGGCACCACGCTGGAAATCAAA (SEQ ID NO: 172)NIVLTQSPASLPVSLGQRATMSCRASKSVDSYGTSFMHWYQHRPGQPPKLLISLASNLESGVPGRFSGSGSRTDFTLTIDPVEPDDAATYYCQQNNEDPRTFGGGTTLEIK (SEQ ID NO: 173) 537G7A6GACATTGTGCTGACACAGTCTCCTGCTTCTTTGGCTGTGTCTGTAGGGCAGAGGGCCACCGTATCCTGCAGAGTCAGTGAAAGTGTTGATAGATATGCCGATAGTTTTATGCACTGGTACCAGCAGAAACCAGGACAGCCACCCAAACTCCTCATCTATCTTGCATCCAACCTAGAATCTGGGGTCCCTGCCAGGTTCAGTGGCAGTGGGTCTAGGACAGACTTCACCCTCACCATTGATCCTGTGGAGGCTGATGATGCTGCAACCTATTACTGTCAGCAAAATAAAGAGGATCCGTACACGTTCGGAGGGGGGACCAAGCTGGAACTTAAA (SEQ ID NO: 174)DIVLTQSPASLAVSVGQRATVSCRVSESVDRYADSFMHWYQQKPGQPPKLLIYLASNLESGVPARFSGSGSRTDFTLTIDPVEADDAATYYCQQNKEDPYTFGGGTKLELK (SEQ ID NO: 175) 551H4D6GACATTGTGCTGACCCAATCTCCAGCTTCTTTGGCTGTGTCTCTAGGGCAGAGGGCCACCATGTCCTGCAGAGCCAGTGAAAGTGTTGATAGTTATGGCAATAGTTTTATACACTGGTACCAGCAGAAACCAGGACAGCCACCCAGACTCCTCATCTATCGTGCATCCAACCTAAATTCTGGGATCCCTGCCAGGTTCAGTGGCAGTGGGTCTAGGACAGACTTCACCCTCACCATTAGTTCTGTGGAGGCTGATGATGTTGCAACCTATTACTGTCACCAAAATAATGAGGATCCTCGGACGTTCGGTGGAGGCACCAAGCTGGAAATCAAA (SEQ ID NO: 176)DIVLTQSPASLAVSLGQRATMSCRASESVDSYGNSFIHWYQQKPGQPPRLLIYRASNLNSGIPARFSGSGSRTDFTLTISSVEADDVATYYCHQNNEDPRTFGGGTKLEIK (SEQ ID NO: 177) 560H2A7GACATTGTGCTGACCCAATCTCCAGCTTCTTTGGCTGTGTCTCTAGGGCAGAGGGCCACCATCTCCTGCAGAGCCAGCGAAAGTATTGATAATTATGGCCTTATTTTTATGAGCTGGTTCCAACAGAAACCAGGACAGCCACCCAAACTCCTCATCTATGCTGCATCCAACCGAGGATCCGGGGTCCCTGCCAGGTTTAGTGGCAGTGGGTCTGGGACAGACTTCAGCCTCAACATCCATCCTATGGAGGAGGATGATACTGCAATGTATTTCTGTCAGCAAAGTAAGGAGGTTCCGTGGACGTTCGGTGGAGGCACCAAGCTGGAAGTCAAA (SEQ ID NO: 178)DIVLTQSPASLAVSLGQRATISCRASESIDNYGLIFMSWFQQKPGQPPKLLIYAASNRGSGVPARFSGSGSGTDFSLNIHPMEEDDTAMYFCQQSKEVPWTFGGGTKLEVK (SEQ ID NO: 179) 606H7F8GACATCCAGATGACTCAGTCTCCAGCCTCCCTATCTGCATCTGTGGGAGAAACTGTCACCATCACATGTCGAGCAAGTGGGAATATTCACAATTATTTAGCATGGTATCAGCAGAAACAGGGAAAATCTCCTCAGCTCCTGGTCTATAATGCAAAAACCTTAGCAGATGGTGTGCCATCAAGGTTCAGTGGCAGTGGATCAGGAACACAATTTTCTCTCAAGATCAACAGCCTGCAGCCTGAAGATTTTGGGAGTTATTACTGTCAACATTTTTGGAGTACTCCGTACACGTTCGGAGGGGGGACCAAGCTGGAAATAAAA (SEQ ID NO: 180)DIQMTQSPASLSASVGETVTITCRASGNIHNYLAWYQQKQGKSPQLLVYNAKTLADGVPSRFSGSGSGTQFSLKINSLQPEDFGSYYCQHFWSTPYTFGGGTKLEIK (SEQ ID NO: 181)

TABLE 2 Anti-BAFF Mouse Leads - VH Sequences Designation Sequence206G9A10 CAGGTCCAACTGCAGCAGCCTGGGGCTGAGCTTGTGAAGCCTGGGGCTTCAGTGAAGCTGTCCTGCAAGGCTTCTGGCTACACCTTCAGTATCTTCTGTATACACTGGGTGCAACAGAGGCCTGGACGAGGCCTTGAGTGGATTGGAAGGATTGATCCTAGTAGTGGTGGTACTAAGTACAATGAGAAGTTCGAGAGCAAGGCCACACTGACTGTAGACAAATCCTCCAGCACAGCCTACATGCAGCTCAGCAGCCTGACATCTGAGGACTCTGCGGTCTATTATTGTGCAAGAGGGGAGGATTTATTAGTACGGACGGATGCTATGGACTACTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCA (SEQ ID NO: 58)QVQLQQPGAELVKPGASVKLSCKASGYTFSIFCIHWVQQRPGRGLEWIGRIDPSSGGTKYNEKFESKATLTVDKSSSTAYMQLSSLTSEDSAVYYCARGEDLLVRTDAMDYWGQGTSVTVSS (SEQ ID NO: 59) 227D5A7CAGGTCCAACTGCAGCAGCCTGGGGCTGAACTTGTGAAGCCTGGGGCTTCAGTGAAGCTGTCCTGCAAGGCTTCTGGCTACACCTTCAGTATTTTCTGTGTACACTGGGTGCAACAGAGGCCTGGACGAGGCCTTGAGTGGATTGGAAGGATTGATCCTAGTAGTGGTGGTACTAAGTACAATGAGAAGTTCGAGAGCAAGGCCACACTGACTGTAGACAAATCGTCCAGCACAGCCTACATGCAGCTCAGCAGCCTGACACCTGAGGACTCTGCGGTCTATTATTGTGCAAGAGGGGAGGATTTATTAGTACGGACGGATGCTCTGGACTACTGGGGTCAAGGATCCTCAGTCACCGTCTCCTCA (SEQ ID NO: 60)QVQLQQPGAELVKPGASVKLSCKASGYTFSIFCVHWVQQRPGRGLEWIGRIDPSSGGTKYNEKFESKATLTVDKSSSTAYMQLSSLTPEDSAVYYCARGEDLLVRTDALDYWGQGSSVTVSS (SEQ ID NO: 61) 250E5A11CAGGTCCAACTGCAGCAGCCTGGGACTGAGCTTGTGAAGCCTGGGGCTTCAGTGAAGCTGTCCTGCAAGGCTTCTGGCTACACCTTCAGTATCTTCTGTATACACTGGGTGCAACAGAGGCCTGGACGAGGCCTTGAGTGGATTGGAAGGATTGATCCTAGTAGTGGTGGCACTAAATATAATGAGAGGTTCGAAAACAAGGCCACACTGACTGTAGACAAATCCTCCAGCACAGCCTACATGCAGCTCAGCAGTCTGACATTTGAGGACTCTGCGGTCTATTATTGTGCAAGAGGGGAGGATTTATTAGTACGGACGGATGCTATGGACTACTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCA (SEQ ID NO: 62)QVQLQQPGTELVKPGASVKLSCKASGYTFSIFCIHWVQQRPGRGLEWIGRIDPSSGGTKYNERFENKATLTVDKSSSTAYMQLSSLTFEDSAVYYCARGEDLLVRTDAMDYWGQGTSVTVSS (SEQ ID NO: 63) 227D3B11CAGGTCCAACTGCAGCAGCCTGGGGCTGAGCTTGTGAAGCCTGGGGCTTCAGTGAAGCTGTCCTGTAAGGCTTCTGGCTACTCCTTCAGCACCTTCTTTATACACTGGATACAGCAGAGGCCTGGGCGAGGCCTTGAGTGGATTGGAAGGATTGATCCTAATAGTGGTGGTACTAAGTACAATGAGAAGTTCGAGAGTAAGGCCACACTGACTGTTGACAAACCCTCCAGTACAGCCTACATGCACCTCAGCAGCCTGACATCTGAGGACTCTGCGGTCTATTATTGTGCAAGAGGGGAGGATTTATTGATACGGACGGATGCTATGGACTACTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCA (SEQ ID NO: 64)QVQLQQPGAELVKPGASVKLSCKASGYSFSTFFIHWIQQRPGRGLEWIGRIDPNSGGTKYNEKFESKATLTVDKPSSTAYMHLSSLTSEDSAVYYCARGEDLLIRTDAMDYWGQGTSVTVSS (SEQ ID NO: 65) 235F5B9CAGGTCCAACTGCAGCAGCCTGGGGCTGAGCTTGTGAAGCCTGGGGCTTCAGTGAAGCTGTCCTGCAAGGCTTCTGGCTACTCCTTCAGTACCTTCTTTATACACTGGATACAGCAGAGGCCTGGGCGAGGCCTTGAGTGGATTGGAAGGATTGATCCTAATAGTGGTGCTACTAAATACAATGAGAAGTTCGAGAGTAAGGCCACACTGACTGTTGACAAACCCTCCAGTACAGCCTACATGCACCTCAGCAGCCTGACATCTGAGGACTCTGCGGTCTATTATTGTGCAAGAGGGGAGGATTTATTGATTCGGACGGATGCTCTGGACTACTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCA (SEQ ID NO: 66)QVQLQQPGAELVKPGASVKLSCKASGYSFSTFFIHWIQQRPGRGLEWIGRIDPNSGATKYNEKFESKATLTVDKPSSTAYMHLSSLTSEDSAVYYCARGEDLLIRTDALDYWGQGTSVTVSS (SEQ ID NO: 67) 217H12A7CAGGTCCAACTGCAGCAGCCTGGGGCTGAGCTTGTGAAGCCTGGGGCTTCAGTGAAGCTGTCCTGCAAGGCTTCTGGCTACACCTTCAGTACCTTCTTAATACACTGGGTGCAGCAGAGGCCTGGACGAGGCCTTGAGTGGATTGGAAGGATTGATCCTAATAGTGGTGGTACTAAGTACAATGAGAAGTTCGAGAGGAAGGCCACACTGACTGTAGACAAACCCTCCAGCACAGCCTACATGCAGCTCAGCAGCCTGACATCTGAGGACTCTGCGGTCTATTATTGTGCAAGAGGGGAGGATTTATTACTACGGACGGATGCTATGGACTACTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCA (SEQ ID NO: 68)QVQLQQPGAELVKPGASVKLSCKASGYTFSTFLIHWVQQRPGRGLEWIGRIDPNSGGTKYNEKFERKVTLTVDKPSSTAYMQLSSLTSEDSAVYYCARGEDLLLRTDAMDYWGQGTSVTVSS (SEQ ID NO: 69) 210D9B8CAGGTCCAACTGCAGCAGCCTGGGACTGAATTTGTGAAGCCTGGGGCTTCAGTGAAGCTGTCCTGCGAGGCTTCTGGCTACACCTTCATCACCTACTGGATGCACTGGGTGAAGCAGAGGCCTGGACGAGGCCTTGAGTGGATTGGAGGGATTGATCCTAATAGTGGTGTTATTAAGTACAATGAGAAGTTCAAGAGTAAGGCCACACTGACTGTAGACAAACCCTCCAGCACAGCCTACATGCAGCTCAGCAGCCTGACATCTGAGGACTCTGCGGTCTACTATTGTGCAAGAGGGGAGGATTTATTAATACGGACGGATGCTATGGACTACTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCA (SEQ ID NO: 70)QVQLQQPGTEFVKPGASVKLSCEASGYTFITYWMHWVKQRPGRGLEWIGGIDPNSGVIKYNEKFKSKATLTVDKPSSTAYMQLSSLTSEDSAVYYCARGEDLLIRTDAMDYWGQGTSVTVSS (SEQ ID NO: 71) 214G4B7CAGGTCCAACTGCAGCAGCCTGGGGCTGAGTTTGTGAAGCCTGGGGCTTCAGTGAAGCTGTCCTGCAAGGCTTCTGGCTACTCCTTCAGTACCTTCTGTATACACTGGGTGCAGCAGAGGCCTGGGCGAGGCCTTGAGTGGATTGGAAGGATTGATCCTAATAGTGGTGGTACTAAATACAATGAGAAGTTCGAGAGTAAGGCCACACTGACTATAGACAAACCCTCCAGTACAGCCTACGTGCACCTCAGCAGCCTGACATCTGAGGACTCTGCGGTCTATTATTGTGCAAGAGGGGAGGATTTATTGATACGGACGGATGCTATGGACTACTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCA (SEQ ID NO: 72)QVQLQQPGAEFVKPGASVKLSCKASGYSFSTFCIHWVQQRPGRGLEWIGRIDPNSGGTKYNEKFESKATLTIDKPSSTAYVHLSSLTSEDSAVYYCARGEDLLIRTDAMDYWGQGTSVTVSS (SEQ ID NO: 73) 13J018-1A4CAGGTTCAGCTGCAGCAGTCTGGACCTGAGGTGGTGAGGCCTGGGGCTTCAGTGAAGATATCCTGCAAGGCTCCTGACCATATTTTCAGTATCCACTGGATGCAGTGGGTAAGACAGAGGCCTGGACCGGGCCTTGAGTGGATTGGAGAGATTTTTCCTGGAAGTGGTACTACTGATTATAATGAGAAATTCAAGGGCAAGGCCACAGTGACGGTAGATAGAGGCTCCAGGTCAGCCTACATGCAGTTCAACAGCCTGACATCTGAGGACTCTGCGGTCTATTTCTGTGCAAGCGGAGCCTTTGACTACTGGGGCCAAGGCACCACTCTCACAGTCTCTT CA (SEQ ID NO: 74)QVQLQQSGPEVVRPGASVKISCKAPDHIFSIHWMQWVRQRPGPGLEWIGEIFPGSGTTDYNEKFKGKATVTVDRGSRSAYMQFNSLTSEDSAVYFCASGAFDYWGQGTTLTVSS (SEQ ID NO: 75) 1002E8A6GAGGTCCAGCTGCAGCAGTCTGGACCTGAGCTGGTAAAGCCTGGGGCTTCAGTGAAGATGTCCTGCAAGGCTTCTGGATACACAATCACTAGTTATGTTATGCACTGGGTGAAGCAGAAGCCTGGGCAGGGCCTTGAGTGGATTGGATATATTAATCCTAACAATGATGGCACTAAGTACAATGAGAAGTTCAAAGGCAAGGCCACACTGACTTCAGACAAATCCTCCAACACAGCCTACATGGAGCTCAGCAGCCTGACCTCTGAGGACTCTGCGGTCTATTATTGTGCAAGAGGGGACTATAGTAACTACTTCTACTGGTACTTCGATGTCTGGGGCGCAGGGACCACGGTCACCGTCTCCTCA (SEQ ID NO: 182)EVQLQQSGPELVKPGASVKMSCKASGYTITSYVMHWVKQKPGQGLEWIGYINPNNDGTKYNEKFKGKATLTSDKSSNTAYMELSSLTSEDSAVYYCARGDYSNYFYWYFDVWGAGTTVTVSS (SEQ ID NO: 183) 1070A6B7CAGGTCCCGCTGCAGCAGCCTGGGGCTGAGATGGTGAGGCCTGGGGCTTCAATGAGGTTGTCCTGTAAGGCTTCTGGCTACACCTTCCCCGGCTACTGGATGCACTGGGTGAAGCAGAGGCCTAGACAAGGCCTTGAGTGGATTGCTAAGATTGATCCCTCTGATAGTGAAACTCACTACAATCAAAACTTCAAGGACAAGGCCACATTGACTGTAGACAAATATTCCAACACAGTCTACATGCAGCTCAACAGCCTGACATCTGAAGACTCTGCGGTCTATTACTGTGCAAACGAGGGTTGGGACAGCCTTACGAAAGTCTGGTTTGGTTGGTGGGGCCAAGGGACTCTGGTCACTGTCTCTGCA (SEQ ID NO: 184)QVPLQQPGAEMVRPGASMRLSCKASGYTFPGYWMHWVKQRPRQGLEWIAKIDPSDSETHYNQNFKDKATLTVDKYSNTVYMQLNSLTSEDSAVYYCANEGWDSLTKVWFGWWGQGTLVTVSA (SEQ ID NO: 185) 1094C4E6GAGGTTCAGCTGCAGCAGTCTGGGGCTGAGCTTGTGAGGCCAGGGGCCTCAGTCAAGTTGTCCTGCACAGCTTCTGGCTTTAACATTAAAGACGACTATATGCACTGGGTGAAGCAGACGCCTGAACAGGGCCTGGAGTGGATTGGAAGGATTGATCCTGCGTATGGTAATGGTAAGTATGTCCCGAAGTTCCAGGACAAGGCCACTATAACTGCAGACACATCCTCCAACACAGCCTACCTGCAGCTCAGCAGCCTGACATCTGAGGACACTGCCGTCTATTACTGTGCTAGACGGTACTACGCTGTTAGTTCCGTAGACTATGCTCTGGACTACTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCA (SEQ ID NO: 186)EVQLQQSGAELVRPGASVKLSCTASGFNIKDDYMHWVKQTPEQGLEWIGRIDPAYGNGKYVPKFQDKATITADTSSNTAYLQLSSLTSEDTAVYYCARRYYAVSSVDYALDYWGQGTSVTVSS (SEQ ID NO: 187) 27I21-3C7CAGGTTCAGCTGCAGCAGTCTGGACCTGAGCTGGTGAGGCCTGGGACTTCAGTGAAGATATCCTGCAAGGCTCCTGGCTATATCTTCACCAGCCACTGGATGCAGTGGGTAAGACAGAGGCCTGGACAGGGCCTTGAGTGGATTGGAGACATTTTTCCTGGAAGCGGTACTACTGATTATAATGAGAAGTTCAAGGACAAGGCCACAGTGACGGTAGACAGATCCTCCAGTTCAGCCTACATGCAGTTCAACAGCCTGACATCTGAGGACTCTGCGGTCTATTTCTGTGCAAGCGGAGCCTTTGACTACTGGGGCCAAGGCACCACTCTCACAGTCTCCTC A (SEQ ID NO: 188)QVQLQQSGPELVRPGTSVKISCKAPGYIFTSHWMQWVRQRPGQGLEWIGDIFPGSGTTDYNEKFKDKATVTVDRSSSSAYMQFNSLTSEDSAVYFCASGAFDYWGQGTTLTVSS (SEQ ID NO: 189) 317H2A6CAGGTTACTCTGAAAGAGTCTGGCCCTGGGATATTGCAGCCCTCCCAGACCCTCAGTCTGACTTGTTCCTTCTCTGGGTTTTCACTGAGGACTTTTGGCATGGGTGTAGGCTGGATTCGTCAGCCTTCAGGGAAGGGTCTGGAGTGGCTGGCACACATTTGGTGGAATGGTGATAAATACTATGACCCAGCCCTGAAGAGTCGGCTCACAATTTCCAAGGATACCTCCGAAAACCGGGTATTCCTCAATATCGCCAATGTGGACACTACAGATACTGCCCCATACTACTGTGTTCGAATTGGTCCTTCTATTACTACGGTAGCAGAGGGATTTGCTTACTGGGGCCAAGGGACTCTGGTCACTGTCTCTGCA (SEQ ID NO: 190)QVTLKESGPGILQPSQTLSLTCSFSGFSLRTFGMGVGWIRQPSGKGLEWLAHIWWNGDKYYDPALKSRLTISKDTSENRVFLNIANVDTTDTAPYYCVRIGPSITTVAEGFAYWGQGTLVTVSA (SEQ ID NO: 191) 319B8A12AAGGTTACTCTGAAAGAGTCTGGCCCTGGGATATTGCAGCCCTCCCAGACCCTCAGTCTGACTTGTTCCTTCTCTGGATTTTCACTGAGGACTTTTGGTATGGGTGTAGGCTGGATTCGTCAGCCTTCAGGGAAGGGTCTGGAGTGGCTGGCACACATTTGGTGGAATGATGAGAAATACTATAATCCAGACCTGAAGAGTCGGCTCACAGTTTCCAAGGATTCCTCCAAAAACCAGGTATTCCTCACGATCGCCAATGTGGACACTTCAGATACTGCCCCATACTACTGTACTCGAGTTGGTCCTTCTATTTCTACGGTTGCAGAGGGATTTCCTTACTGGGGCCAAGGGACTCTGGTCACTGTCTCTGCA (SEQ ID NO: 192)KVTLKESGPGILQPSQTLSLTCSFSGFSLRTFGMGVGWIRQPSGKGLEWLAHIWWNDEKYYNPDLKSRLTVSKDSSKNQVFLTIANVDTSDTAPYYCTRVGPSISTVAEGFPYWGQGTLVTVSA (SEQ ID NO: 193) 320F9C5CAGGTTACTCTGAAAGAGTCTGGCCCTGGGATATTGCAGTCCTCCCAGACCCTCAGTCTGACTTGTTCTTTCTCTGGGTTTTCACTGAGGACCTTTGGTATGGGTGTAGGCTGGATTCGTCAACCTTCAGGGAAGGGTCTGGAATGGCTGGCACACATTTGGTGGAATGATGATAAGTCCTCTCACCCAGCCCTGAAGAGTCGTCTCACAATCTCCAAGGATACCTCCAAAAACCAGGTATTCCTCAAGATCGCCAATGTGGACACTGCAGAAACTGCCACATATTATTGTGTTCGAATAGGTCCTTCAATTACTACGGTTGCAGAGGGGTTTGCTTACTGGGGCCAAGGGACTCTGGTCACTGTCTCTGCA (SEQ ID NO: 194)QVTLKESGPGILQSSQTLSLTCSFSGFSLRTFGMGVGWIRQPSGKGLEWLAHIWWNDDKSSHPALKSRLTISKDTSKNQVFLKIANVDTAETATYYCVRIGPSITTVAEGFAYWGQGTLVTVSA (SEQ ID NO: 195) 323E9D1CAGGTTACTCTGAAAGAGTCTGGCCCTGGGATATTGCAGCCCTCCCAGACCCTCAGTCTGACTTGTTCCTTCTCTGGATTTTCAATGAGGACTTTTGGTATGGGTGTAGGCTGGATTCGTCAGCCTTCAGGGAAGGGTCTGGAGTGGCTGGCACACATTTGGTGGAATGATGAGAAATACTATAATCCAGACCTGAAGAGTCGGCTCACAGTTTCCAAGGATTCCTCCAAAAACCAGGTATTCCTCACGATCGCCAATGTGGACACTTCAGATACTGCCCCATACTACTGTACTCGAGTTGGTCCTTCTATTTCTACGATTGCAGAGGGATTTCCTTACTGGGGCCAAGGGACTCTGGTCACTGTCTCTGCA (SEQ ID NO: 196)QVTLKESGPGILQPSQTLSLTCSFSGFSMRTFGMGVGWIRQPSGKGLEWLAHIWWNDEKYYNPDLKSRLTVSKDSSKNQVFLTIANVDTSDTAPYYCTRVGPSISTIAEGFPYWGQGTLVTVSA (SEQ ID NO: 197) 332C1B12CAGGTCCAACTGCAGCAGCCTGGGGCTGAATTGGTGAAGCCTGGGGCTTCAGTGAAGCTGTCCTGCAAGGCTTCTGGCTACACTTTCACCAACGACAATTACTGGATGAACTGGATGAAACAGAGGCCTGGACGAGGCCTCGAGTGGATTGGAAGGATTCGTCCTTCTGATAGTGAAACTCACTACAATCAAAAATTCACGAACAAGGCCACACTGACTGTAGACAAATCCTCCAGCACAGCCTACATCCAACTCAGCAGCCTGACATCTGTGGACTCTGCGGTCTATTATTGTGCAAGATCTTGGGAAGATTTATTACTACGATCGATGGAGGACTACTTTGACTACTGGGGCCAAGGCACCACTCTCACAGTCTCCTCA (SEQ ID NO: 198)QVQLQQPGAELVKPGASVKLSCKASGYTFTNDNYWMNWMKQRPGRGLEWIGRIRPSDSETHYNQKFTNKATLTVDKSSSTAYIQLSSLTSVDSAVYYCARSWEDLLLRSMEDYFDYWGQGTTLTVSS (SEQ ID NO: 199) 344B9D9GAGTTCCAACTGCAGCAGTCTGGACCTGAGCTGGGGGAGCCTGGCGCTTCAGTGAAAATCTCCTGCAAGGCTTCTGGTTTCTCATTCAGTGACTACAACATAAATTGGGTGAAGCAGAGCAATGGAAAGAGTCTTGAGTGGATTGGAAAAGTTCATCCTAAGGATGGTACTGCTACCTACAATCAGAAGTTCCAGGACAAGGCCACATTGACTCTAGACCAGTCTTCCAGCACAGCCTACATGCAACTCAGCAGCCTGACATCGGAGGACTCTGCAGTCTATTACTGTCTCCCGCTCTACTATGATTCCCTGACAAAAATTTTGTTTGCTTATTGGGGCCAAGGGACTCTGGTCACTGTCTCTGCA (SEQ ID NO: 200)EFQLQQSGPELGEPGASVKISCKASGFSFSDYNINWVKQSNGKSLEWIGKVHPKDGTATYNQKFQDKATLTLDQSSSTAYMQLSSLTSEDSAVYYCLPLYYDSLTKILFAYWGQGTLVTVSA (SEQ ID NO: 201) 348A6C1CAGGTTACTCTGAGAGAGTCTGGGCCTGGGATATTGCAGCCCTCCCAGACCCTCAGTCTGACTTGTTCCTTCTCTGGGTTTTCACTGAGGACCTTTGGTATGGGTGTAGGCTGGATTCGTCAGCCTTCAGGGAAGGGTCTGGAGTGGCTGGCACACATCTGGTGGAATGATGAGAAATATTATAACCCAGCCCTGAAGAGTCGGCTCACAGTTTCCAAGGATTCCTCCGAAAACCAGGTATTCCTCAAGATCGCCAATGTGGACACTACAGATACTGCCCCATACTACTGTGCTCGACTTGGTCCTTCTATTACTACGGTTGCAGAGGGATTTCCGTACTGGGGCCAAGGGACTCTGGTCACTGTCTCTGCA (SEQ ID NO: 202)QVTLRESGPGILQPSQTLSLTCSFSGFSLRTFGMGVGWIRQPSGKGLEWLAHIWWNDEKYYNPALKSRLTVSKDSSENQVFLKIANVDTTDTAPYYCARLGPSITTVAEGFPYWGQGTLVTVSA (SEQ ID NO: 203) 352G11A10CAGGTCCAGCTGCAGCAGTCTGGACCTGAGCTGGTGAAGCCCGGGGGTTCAGTGAGGATATCCTGCAAGGCTTCTGGTTACAGCCTCATAAGCTACTATATACACTGGGTGAAACAGAGGCCGGGACAGGGCCTTGAGTGGATTGGATTGACTTTTCCTGGAAGTGGTAATTCTAAGTTCATTGAGAAGTTCAAGGGCAAGGCCACACTGACGGCAGACACATCCTCCAACACTGCCTACATACAGCTCAGCAGTCTAACATCTGAGGACTCTGCGGTCTATTACTGTACAAGGGGGGACTTCGGTAACTACCTTGCCTACTGGTACTTCGATGTCTGGGGCACAGGGACCACGGTCACCGTCTCCTCA (SEQ ID NO: 204)QVQLQQSGPELVKPGGSVRISCKASGYSLISYYIHWVKQRPGQGLEWIGLTFPGSGNSKFIEKFKGKATLTADTSSNTAYIQLSSLTSEDSAVYYCTRGDFGNYLAYWYFDVWGTGTTVTVSS (SEQ ID NO: 205) 363D4A10CAGGTTACTCTGAAAGAGTCTGGCCCTGGGATATTGCAGTCCTCCCAGACCCTCAGTCTGACTTGTTCTTTCTCTGGGTTTTCACTGAAGACCTTTGGTATGGGTGTGGGCTGGATTCGTCAGCCTTCAGGGAAGGGTCTGGAGTGGCTGGCACACATTTGGTGGAATGATGATAAATTCTATCACCCAGCCCTGAAGAGTCGGCTCACAATCTCCAAGGATACCTCCAAAAACCAGGTATTCCTCAAGATCGCCAATGTGGACACTGCAGAAACTGCCACATACTACTGTGTTCGAATTGGTCCTTCAATTACTACGGTAGCAGAGGGGTTTGCTTACTGGGGCCAAGGGACTCTGGTCACTGTCTCTGCA (SEQ ID NO: 206)QVTLKESGPGILQSSQTLSLTCSFSGFSLKTFGMGVGWIRQPSGKGLEWLAHIWWNDDKFYHPALKSRLTISKDTSKNQVFLKIANVDTAETATYYCVRIGPSITTVAEGFAYWGQGTLVTVSA (SEQ ID NO: 207) 381A6A9CAGGTCCAGCTGCAGCAGTCTGGACCTGAGCTGGTGAAGCCCGGGGGTTCAGTGAAGATATCCTGCAAGGCTGCTGGCTACAGCCTCACAAGCTACTATATACACTGGGTGAAGCAGAGGCCGGGACAGGGACTTGAGTGGATTGGATTGATTTTTCCTGGAAGTGGTAATTCTAAGTACATTGAGAAGTTCAAGGGCAAGGCCACACTGACGGCGGACACATCCTCCAACACTGCCTACATGCAGCTCAGCAGCCTAACATCTGAGGACTCTGCGGTCTATTATTGTACAAGGGGGGACTTCGGTAACTACCTTGCCTACTGGTACTTCGATGTCTGGGGCACAGGGACCACGGTCACCGTCTCCTCA (SEQ ID NO: 208)QVQLQQSGPELVKPGGSVKISCKAAGYSLTSYYIHWVKQRPGQGLEWIGLIFPGSGNSKYIEKFKGKATLTADTSSNTAYMQLSSLTSEDSAVYYCTRGDFGNYLAYWYFDVWGTGTTVTVSS (SEQ ID NO: 209) 384D5A2CAGGTCACTCTGAAAGAGTCTGGCCCTGGAATATTGCAGCCCTCCCAGACCCTCAGTCTGACTTGTTCTTTCTCTGGGTTTTCACTGAACACTTATGGTATGGGTGTGGGTTGGATTCGTCAGCCTTCAGGGAAGGGTCTGGAGTGGCTGGCCAACATTTGGTGGAATGATGATAAGTACTATAACTCAGCCCTGAAGAGCCGGCTCGCAATCTCCAAAGATGCCTCCAACAGCCAGGTATTCCTCAAGATCTCCAGTGTGGACACTACAGATACTGCCACATACTACTGTGCTCAAGTAGCCGCTACTATAGTAACTACGTACGGGGCCTGGTTTGCTTACTGGGGCCAAGGGACTCTGGTCACTGTCTCTGCA (SEQ ID NO: 210)QVTLKESGPGILQPSQTLSLTCSFSGFSLNTYGMGVGWIRQPSGKGLEWLANIWWNDDKYYNSALKSRLAISKDASNSQVFLKISSVDTTDTATYYCAQVAATIVTTYGAWFAYWGQGTLVTVSA (SEQ ID NO: 211) 394F5A5GTTACTCTGAAAGAGTCTGGCCCTGGGATATTGCAGCCCTCCCAGACCCTCAGTCTGACTTGTTCCTTCTCTGGGTTTTCACTGAGGACTTTTGGTATGGGTGTAGGCTGGATTCGTCAGCCTTCAGGGAAGGGTCTGGAGTGGCTGGCACACATTTGGTGGAATGATGAGAAATATTATAATCCAACCCTGAAGAGTCGGCTCACAATTTCCAAGGATACCTCCAAAAACCAGGTATTCCTCAGGATCGCCAATGTGGACACTGCAGTTACTGCCGCATACTACTGTGCTCGAATAGGTCCTTCTATTACTACGGTAGTAGAGGGATTTCCTTACTGGGGCCAAGGGACTCTGGTCACTGTCTCTGCA (SEQ ID NO: 212)VTLKESGPGILQPSQTLS LTCSFSGFSLRTFGMGVGWIRQPSGKGLEWLAHIWWNDEKYYNPTLKSRLTISKDTSKNQVFLRIANVDTAVTAAYYCARIGPSITTVVEGFPYWGQGTLVTVS A (SEQ ID NO: 213) 409F12A11ATCCAGCTGCAGCAGTCTGGACCTGAGCTGGTGAAGCCTGGGGCTTCAATGAAGATATCGTGCAAGGCTTCTGGCTACACCTTCACTGACAAGTATATAAACTGGGTGAAGCAGAGGCCTGGACAGGGACTTGAGTGGATTGGATGGATTTATCCTGGAAGCGGTAATACTAAGTACAATGAGAAGTTCAAGGGCATGGCCACATTGACTGTAGACACATCCTCCAATACAGCCTATATACATCTCAGCAGCCTGACCTCTGAGGACTCTGCGGTCTATTTCTGTGCACGAGGAATTATTTATTACTACGATGGTTCATACCCCTATGCTTTGGACTACTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCA (SEQ ID NO: 214)IQLQQSGPELVKPGASMKISCKASGYTFTDKYINWVKQRPGQGLEWIGWIYPGSGNTKYNEKFKGMATLTVDTSSNTAYIHLSSLTSEDSAVYFCARGIIYYYDGSYPYALDYWGQGTSVTVSS (SEQ ID NO: 215) 418F6D9CAGGTCCAGCTGCAGCAGTCTGGACCTGAACTGGTGAAGCCTGGAGCTTCAGTGAAGTTGTCCTGCAAGGCTTCTGGCTACACCTTCACTGACTATAGTATACACTGGGTGAAGCAGAGTCCTGGACAGGGACTTGAGTGGATTGGATGGATTTATCCTGGAAGTGGTAATACTAAGTACAATGACAAGTTCAAGGGCAAGGCCACAATGACTGCAGACAAATCCTCCAGAACAGTCTACATGCAGCTCAGCAGCCTGACGTCTGAGGAGTCTGCGGTCTATTTCTGTGCAAGAGACTACCGGCGATACTATGCTATAGACTACTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCA (SEQ ID NO: 216)QVQLQQSGPELVKPGASVKLSCKASGYTFTDYSIHWVKQSPGQGLEWIGWIYPGSGNTKYNDKFKGKATMTADKSSRTVYMQLSSLTSEESAVYFCARDYRRYYAIDYWGQGTSVTVSS (SEQ ID NO: 217) 431G5A3CAGCTGCAGCAGTCTGGACCTGAGCTGGTGAAGCCTGGAGCTTCAGTGAAGCTGTCCTGCAAGGCTTCTGGCTACACCTTCACTGACTATAGTATACACTGGGTGAAACAGAGTCCTGGACAGGGACTTGAGTGGATTGGATGGATTTATCCTGGAAGTGATAATACTAAGTACAATGACAAGTTCAAGGGCAAGGCCTCAATGACTGCAGACAAATCCTCCAGAACAGTCTACATGCACCTCAGCAGCCTGACGTCTGAGGAATCTGCGGTCTATTTCTGTGCAAGAGACTACCGGCGGTACTATGCTATGGACTACTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCA (SEQ ID NO: 218)QLQQSGPELVKPGASVKLSCKASGYTFTDYSIHWVKQSPGQGLEWIGWIYPGSDNTKYNDKFKGKASMTADKSSRTVYMHLSSLTSEESAVYFCARDYRRYYAMDYWGQGTSVTVSS (SEQ ID NO: 219) 435A6B3GAGGTCCAGCTGCAGCAGTCTGGACCTGAGCTGGTAAAGCCTGGGGCTTCAATGAAGATGTCCTGCAAGGCTTCTGGATACACATTCACTAGCTATGTTATGCACTGGATGAAGCAGAAGCCTGGGCAGGGCCTTGAGTGGATTGGATATCTTAATCCTAACAATGATGGTACTAAGTACAATGAGAAGTTCAAAGGCAAGGCCACACTGACTTCAGACAAATCCTCCAGCACAGCCTACATGGAGCTCAGCAGCCTGACCTCTGAGGACTCTGCGGTCTATTACTGTGCAAGGGGGGACTATAGTAATTACTTCTACTGGTACTTCGATGTCTGGGGCGCAGGGACCACGGTCTCCGTCTCCTCA (SEQ ID NO: 220)EVQLQQSGPELVKPGASMKMSCKASGYTFTSYVMHWMKQKPGQGLEWIGYLNPNNDGTKYNEKFKGKATLTSDKSSSTAYMELSSLTSEDSAVYYCARGDYSNYFYWYFDVWGAGTTVSVSS (SEQ ID NO: 221) 436H2C12CAGGTCCAGCTGCAGCAGTCTGGACCTGAGCTGGTGAAGCCTGGAGCTTCAGTGAAGCTGTCCTGCAAGGCTTCTGGCTACACCTTCAGTGACTATACTATACACTGGGTGAAGCAGAGTCCTGGACAGGGACTTGAGTGGATTGGATGGATTTACCCTGGAAGGGGTAATACTAAGTACAATGACAAGTTCAAGGGCAAGGCCACAATGACTGCTGACAAATCCTCCAGCACAGCCTACATGCAGCTCAGCAGCCTGACGTCTGAGGAATCTGCGGTCTATTTCTGTGCAAGAGACTACCGGCGGTACTATGCTATGGACTACTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCA (SEQ ID NO: 222)QVQLQQSGPELVKPGASVKLSCKASGYTFSDYTIHWVKQSPGQGLEWIGWIYPGRGNTKYNDKFKGKATMTADKSSSTAYMQLSSLTSEESAVYFCARDYRRYYAMDYWGQGTSVTVSS (SEQ ID NO: 223) 436H6A9CAGCTGCAGCAGTCTGGACCTGAGCTGGTGAAGCCTGGGGCTTCAGTGAAGATATCCTGCAAGGCTTCTGGCTACACCTTCACTGACAACTTTATAAACTGGGTGAAACAGAGGCCTGGACAGGGACTTGAGTGGATTGGATGGATTTCTCCTGGAAGCGGTAATACTAAGAACAATGAGAAGTTCAAGGGCAAGGCCACAGTGACTGTAGACACATCCTCCAGCACAGCCTACATGCAGCTCAGCAGCCTGACCTCTGAGGACTCTGCGGTCTATTTCTGTGCACGAGGAATTATTTATTATTATGATGGTACCTACCCCTATGCTCTGGACTACTGGGGTCAGGGAACCTCAGTCACCGTCTCCTCA (SEQ ID NO: 224)QLQQSGPELVKPGASVKISCKASGYTFTDNFINWVKQRPGQGLEWIGWISPGSGNTKNNEKFKGKATVTVDTSSSTAYMQLSSLTSEDSAVYFCARGIIYYYDGTYPYALDYWGQGTSVTVSS (SEQ ID NO: 225) 440E9D12CAGGTGCGGCTGAGGGAGTCAGGACCTGGCCTGGTGGCGCCCTCCCAGAACCTGTTCATCACATGCACCGTCTCAGGTTTCTCATTAACTGACTATGAAATAAACTGGGTTCGCCAGCCTCCAGGAAAGAATCTGGAGTGGCTGGGAGTGATTTGGACTGGTGGAGGCACAAAATATAATTCAGTTCTCATATCCAGACTGAACATCAGCAAAGACAATTCCAAGAGACAAGTTTTCTTTAAAATGACCAGTCTCCAGACTGATGACACAGCCATATATTACTGTGTAAGAGAGGGGAGGAGATACTATGCTATGGACTACTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCA (SEQ ID NO: 226)QVRLRESGPGLVAPSQNLFITCTVSGFSLTDYEINWVRQPPGKNLEWLGVIWTGGGTKYNSVLISRLNISKDNSKRQVFFKMTSLQTDDTAIYYCVREGRRYYAMDYWGQGTSVTVSS (SEQ ID NO: 227) 441E6F2CGGGTGCAGCTGAAGGAGTCAGGACCTGGCCTGGTGGCGCCCTCACAGAGCCTGTTCATCACATGCACCGTCTCAGGGTTCTCATTAACCACCTATGAAATAAACTGGGTTCGCCAGTCTCCAGGAAAGGGTCTGGAGTGGCTGGGAGTGATATGGACTGGTGGAACCACAAAATATAATTCAGCTTTCATATCCAGACTGAGCATCACCAAAGACAACTCCAAGAGCCTCGTTTTCTTAAAAATGAGCAGTCTGCAAACTGATGACACAGCCATATATTACTGTGTAAGAGAGGGGAGGAGGTACTATGCTATGGACTACTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCA (SEQ ID NO: 228)RVQLKESGPGLVAPSQSLFITCTVSGFSLTTYEINWVRQSPGKGLEWLGVIWTGGTTKYNSAFISRLSITKDNSKSLVFLKMSSLQTDDTAIYYCVREGRRYYAMDYWGQGTSVTVSS (SEQ ID NO: 229) 443C11A12GAGGTCCAGCTGCAGCAGTCTGGACCTGAGCTGGTAAAGCCTGGGGCTTCAGTGAAGATGTCCTGCACGGCTTCTGGATACACATTCACTAGCTATGTTATACACTGGATGAAGCAGAAGCCTGGGCAGGGCCTTGAGTGGATTGGATATCTTCATCGTAACAATGATGGTACTAAGTACAATGAGAAGTTCAAAGTCAAGGCCACACTGACTTCAGACGAATCCTCCAACACAGCCTACATGGAACTCAGCAGCCTGACCTCTGAGGACTCTGCGGTCTATTACTGTGCAAGGGGGGACTATAGTAATTACTTCTACTGGTACTTCGATGTCTGGGGCGCAGGGACTACGGTCTCCGTCTCCTCA (SEQ ID NO: 230)EVQLQQSGPELVKPGASVKMSCTASGYTFTSYVIHWMKQKPGQGLEWIGYLHRNNDGTKYNEKFKVKATLTSDESSNTAYMELSSLTSEDSAVYYCARGDYSNYFYWYFDVWGAGTTVSVSS (SEQ ID NO: 231) 444G1A10CGGGTGCAGCTGAAGGAGTCAGGACCTGGCCTGGTGGCGCCCTCACAGAGCCTGTTCATCACATGCACCGTCTCAGGGTTTTCATTAACCACCTATGAAATAAACTGGGTTCGCCAGTCTCCAGGAAAGGGTCTGGAGTGGCTGGGAGTGATATGGACTGGTGGAACCACAAAATATAATTCAGCTTTCATATCCAGACTGAGCATCACCAAAGACAACTCCAAGAGCCTCGTTTTCTTAAAAATGAGCAGTCTGCAAACTGATGACACAGCCATATATTACTGTGTAAGAGAGGGGAGGAGGTACTATGCTATGGACTACTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCA (SEQ ID NO: 232)RVQLKESGPGLVAPSQSLFITCTVSGFSLTTYEINWVRQSPGKGLEWLGVIWTGGTTKYNSAFISRLSITKDNSKSLVFLKMSSLQTDDTAIYYCVREGRRYYAMDYWGQGTSVTVSS (SEQ ID NO: 233) 450A2A7CAGGTCCAGCTGCAGCAGTCTGGACCTGAGCTGGTGAAGCCTGGAGCTTCAGTGAAGCTGTCCTGCAAGGCTTCTGGCTACACCTTCACTGACTATAGTATACACTGGGTGAAACAGAGTCCTGGACAGGGACTTGAGTGGATTGGATGGATTTATCCTGGAAGTGATAATACTAAGTACAATGACAAGTTCAAGGGCAAGGCCTCAATGACTGCAGACAAATCCTCCAGAACAGTCTACATGCACCTCAGCAGCCTGACGTCTGAGGAATCTGCGGTCTATTTCTGTGCAAGAGACTACCGGCGGTACTATGCTATGGACTACTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCA (SEQ ID NO: 236)QVQLQQSGPELVKPGASVKLSCKASGYTFTDYSIHWVKQSPGQGLEWIGWIYPGSDNTKYNDKFKGKASMTADKSSRTVYMHLSSLTSEESAVYFCARDYRRYYAMDYWGQGTSVTVSS (SEQ ID NO: 235) 456H11B7aCAGGTGCGGCTGAGGGAGTCAGGACCTGGCCTGGTGGCGCCCTCCCAGAACCTGTTCATCACATGCACCGTCTCAGGTTTCTCATTAACTGACTATGAAATAAACTGGGTTCGCCAGCCTCCAGGAAAGAATCTGGAGTGGCTGGGAGTGATTTGGACTGGTGGAGGCACAAAATATAATTCAGTTCTCATATCCAGACTGAACATCAGCAAAGACAATTCCAAGAGACAAGTTTTCTTTAAAATGACCAGTCTCCAGACTGATGACACAGCCATATATTACTGTGTAAGAGAGGGGAGGAGATACTATGCTATGGACTACTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCA (SEQ ID NO: 237)QVRLRESGPGLVAPSQNLFITCTVSGFSLTDYEINWVRQPPGKNLEWLGVIWTGGGTKYNSVLISRLNISKDNSKRQVFFKMTSLQTDDTAIYYCVREGRRYYAMDYWGQGTSVTVSS (SEQ ID NO: 238) 456H11B7bCAGGTTACTCTGAAAGAGTCTGGCCCTGGGATATTGCAGCCCTCCCAGACCCTCAGTCTGACTTGTTCTTTCTCTGGGTTTTCACTGAGCACTTTTGGTATGGGTGTAGGCTGGATTCGTCAGCCTTCAGGGAAGGGTCTGGAGTGGCTGGCACACATTTGGTGGGATGATGATAAGTACTATAACCCAGCCCTGAAGAGTCGGCTCACAATCTCCAAGGATACCTCCAAAAACCAGGTATTCCTCAAGATCGCCAATGTGGACACTGCAGATACTGCCACATACTACTGTGCTCGAATAGAGGGCCCCTACTACTGGTACTTCGATGTCTGGGGCACAGGGACCACGGTCACCGTCTCCTCA (SEQ ID NO: 239)QVTLKESGPGILQPSQTLSLTCSFSGFSLSTFGMGVGWIRQPSGKGLEWLAHIWWDDDKYYNPALKSRLTISKDTSKNQVFLKIANVDTADTATYYCARIEGPYYWYFDVWGTGTTVTVSS (SEQ ID NO: 240) 537G7A6CAGGTGCAGCTGAAGGAGTCAGGACCTGGCCTGGTGGCACCCTCACAGAGCCTGTCCATCACATGCACGGTCTCTGGTTTCTCATTATCCAGATATAGTGTACACTGGATTCGTCAGTCTCCAGGAAAGGGTCTGGAGTGGCTGGGAATGATATGGGGGGGTGGAAACACAGACTACAATTCAGGTCTCAAATCCAGACTGAGCATCAGCAAGGACAACTCCAAGAGCCAAGTTTTCTTAAAAATGAACAGTCTGGAAAATGATGACACAGCCATGTATTACTGTGCCAGCCCCTCCCTCTATTATTATGATGTTGCCTGGTTTCCTTACTGGGGCCAAGGGACTCTGGTCACTGTCTCTGCA (SEQ ID NO: 241)QVQLKESGPGLVAPSQSLSITCTVSGFSLSRYSVHWIRQSPGKGLEWLGMIWGGGNTDYNSGLKSRLSISKDNSKSQVFLKMNSLENDDTAMYYCASPSLYYYDVAWFPYWGQGTLVTVSA (SEQ ID NO: 242) 551H4D6GAGGTTCAGCTGCAGCAGTCTGGGGCTGAGCTTGTGAGGCCAGGGGCCTCAGTCGAGTTGTCCTGCACAGCTTCTGGCTTTAATATTAAAAACGACTATTTGCACTGGGTGAAGCAGAGGCCTGAACAGGGCCTGGAATGGATTGGATGGATTGATTCCGCGAATGATAAGACTAAGTATGCCCCGAAGTTCCAGGACAAGGCCACTATAACTGCAGACCCATCCTCCAACACAGCCTACCTGCAGCTCAGCAGCCTGACATCTGAGGACACTGCCGTCTATTACTGTACTAGAGTTGGGGTTCAGGATGGTTACTACGTTAGGGACTTTGACTACTGGGGCCAGGGCACCACTCTCACAGTCTCCTCA (SEQ ID NO: 243)EVQLQQSGAELVRPGASVELSCTASGFNIKNDYLHWVKQRPEQGLEWIGWIDSANDKTKYAPKFQDKATITADPSSNTAYLQLSSLTSEDTAVYYCTRVGVQDGYYVRDFDYWGQGTTLTVSS (SEQ ID NO: 244) 560H2A7GAGGTGAAGCTGGTGGAGTCTGGAGGAGGCTTGGTACAGCCTGGGGGTTCTCTGAGTCTCTCCTGTGCAGGTTCTGGATTCACCTTCAGTGATTACTACATGAGCTGGGTCCGCCAGCCTCCAGGGAAGGCACTTGAGTGGTTGGCTTTGATTAGAAACAAAGCTCCTGGTTACACAACAGAATACAGTGCATCTGTGAAGGGTCGTTTCACCATCTCCAGAGATAATTCCCAAAGCATCCTCTATCTTCAAATGAATGCCCTGAGACCTGAGGACAGTGCCACTTATTACTGTGCAAGAGTCTTACGACGGGCAGACTGCTTAGACTACTGGGGCCAAGGCACCGCTCTCACAGTCTCCTCA (SEQ ID NO: 245)EVKLVESGGGLVQPGGSLSLSCAGSGFTFSDYYMSWVRQPPGKALEWLALIRNKAPGYTTEYSASVKGRFTISRDNSQSILYLQMNALRPEDSATYYCARVLRRADCLDYWGQGTALTVSS (SEQ ID NO: 246) 606H7F8GTGCAGCTGGTGGAGTCTGGGGGAGGCTTAGTGAAGCCTGGAGGGTCCCTGAAACTCTCCTGTGCAGCCTCTGGATTCACTTTCAGTAGTTATGACATGTCTTGGGTTCGCCAGACTCCAGAGAAGAGGCTGGAGTGGGTCGCAGCCATTAATAGTTATGGTGTTAACACCTACTATCCAGACACTGTGAAGGACCGATTCACCATCTCCAGAGACAATGCCAAGAACACCCTGTACCTGCAAATGAGCAGTCTGAGGTCTGAGGACACAGCCTTGTATTACTGTGCAAGACTTTTAATTGGGCCTTATTACTATGCTATGGACTACTGGGGTCAAGGAACCTCAGTCACCGTCTCCTCA (SEQ ID NO: 247)VQLVESGGGLVKPGGSLKLSCAASGFTFSSYDMSWVRQTPEKRLEWVAAINSYGVNTYYPDTVKDRFTISRDNAKNTLYLQMSSLRSEDTALYYCARLLIGPYYYAMDYWGQGTSVTVSS (SEQ ID NO: 248)

TABLE 3 VκCDR Sequences Designation Sequence VLCDR1 9A10KASQNAGAAVA (SEQ ID NO: 1) VLCDR2 9A10 SASNRYT (SEQ ID NO: 2)VLCDR3 9A10 QQYRSYPRT (SEQ ID NO: 3) VLCDR1 5A7KASQNAGAAVA (SEQ ID NO: 1) VLCDR2 5A7 SASNRYT (SEQ ID NO: 2) VLCDR3 5A7QQYRSFPRT (SEQ ID NO: 4) VLCDR1 5A11 KASQNAGAAVA (SEQ ID NO: 1)VLCDR2 5A11 SASNRYT (SEQ ID NO: 2) VLCDR3 5A11 QQYRSFPRT (SEQ ID NO: 4)VLCDR1 3B11 KASQNAGIDVA (SEQ ID NO: 5) VLCDR2 3B11STSNRYT (SEQ ID NO: 6) VLCDR3 3B11 LQYRSYPRT (SEQ ID NO: 7) VLCDR1 5B9KASQNAGIDVA (SEQ ID NO: 5) VLCDR2 5B9 SKSNRYT (SEQ ID NO: 8) VLCDR3 5B9LQYRSYPRT (SEQ ID NO: 9) VLCDR1 12A7 KASQNAGTAVA (SEQ ID NO: 10)VLCDR2 12A7 SAFNRYT (SEQ ID NO: 11) VLCDR3 12A7QQYRSYPRT (SEQ ID NO: 12) VLCDR1 9B8 KASQSVGIAVA (SEQ ID NO: 13)VLCDR2 9B8 STSNRYT (SEQ ID NO: 6) VLCDR3 9B8 QQYSRYPRT (SEQ ID NO: 14)VLCDR1 4B7 KASQNAGTAVA (SEQ ID NO: 10) VLCDR2 4B7 STSNRYT (SEQ ID NO: 6)VLCDR3 4B7 LQYRSYPRT (SEQ ID NO: 7) VLCDR1 1A4RASQDIGNRLN (SEQ ID NO: 15) VLCDR2 1A4 ATSSLDS (SEQ ID NO: 16)VLCDR3 1A4 LQYASSPFT (SEQ ID NO: 17) VLCDR1 c04RASQDIGNRLS (SEQ ID NO: 76) VLCDR2 c04 ATSSLDS (SEQ ID NO: 16)VLCDR3 c04 LQYASSPFT (SEQ ID NO: 17) VLCDR1 c68RASQDIGNRLH (SEQ ID NO: 77) VLCDR2 c68 ATSSLDS (SEQ ID NO: 16)VLCDR3 c68 LQYASSPFT (SEQ ID NO: 17) VLCDR1 c44RASQDIGNRLP (SEQ ID NO: 78) VLCDR2 c44 ATSSLDS (SEQ ID NO: 16)VLCDR3 c44 LQYASSPFT (SEQ ID NO: 17) VLCDR1 c03RASQDIGNRLR (SEQ ID NO: 79) VLCDR2 c03 ATSSLDS (SEQ ID NO: 16)VLCDR3 c03 LQYASSPFT (SEQ ID NO: 17) VLCDR1 c10RASQDIGNRLM (SEQ ID NO: 80) VLCDR2 c10 ATSSLDS (SEQ ID NO: 16)VLCDR3 c10 LQYASSPFT (SEQ ID NO: 17) VLCDR1 c10.1RASESVDSYGNIFMH (SEQ ID NO: 249) VLCDR2 c10.1 LASNLES (SEQ ID NO: 276)VLCDR3 c10.1 QQNNEAPWT (SEQ ID NO: 293) VLCDR1 c10.2RASKSVSTSGYSYMH (SEQ ID NO: 250) VLCDR2 c10.2 LASNLES (SEQ ID NO: 276)VLCDR3 c10.2 QQNNEAPWT (SEQ ID NO: 293) VLCDR1 8A6KASQDINSYLT (SEQ ID NO: 251) VLCDR2 8A6 RANRLVS (SEQ ID NO: 277)VLCDR3 8A6 LQYDEFPYT (SEQ ID NO: 294) VLCDR1 6B7RCSQSLVHSNGNTYLH (SEQ ID NO: 252) VLCDR2 6B7 KVSDRFS (SEQ ID NO: 278)VLCDR3 6B7 SQSTHVPLT (SEQ ID NO: 295) VLCDR1 4E6KASQDVATAVA (SEQ ID NO: 253) VLCDR2 4E6 WASTRHT (SEQ ID NO: 279)VLCDR3 4E6 QQYSNYPYT (SEQ ID NO: 296) VLCDR1 3C7RASQDIGNRLN (SEQ ID NO: 15) VLCDR2 3C7 ATSSLDS (SEQ ID NO: 16)VLCDR3 3C7 LQYASYPFT (SEQ ID NO: 297) VLCDR1 2A6KASQNVGSAVV (SEQ ID NO: 254) VLCDR2 2A6 SASNRYS (SEQ ID NO: 280)VLCDR3 2A6 QQYSNYPLT (SEQ ID NO: 298) VLCDR1 8A12KASQNVGAAVV (SEQ ID NO: 255) VLCDR2 8A12 SASNRYI (SEQ ID NO: 281)VLCDR3 8A12 QQYSNYPLT (SEQ ID NO: 298) VLCDR1 9C5KASQNVGSVVA (SEQ ID NO: 256) VLCDR2 9C5 SASNRYT (SEQ ID NO: 2)VLCDR3 9C5 QQYSSYPLT (SEQ ID NO: 299) VLCDR1 1B12RASKGVSTSSYTFIH (SEQ ID NO: 257) VLCDR2 1B12 YASNLES (SEQ ID NO: 282)VLCDR3 1B12 QHSREFPRT (SEQ ID NO: 300) VLCDR1 9D9RASGNIHNYLA (SEQ ID NO: 258) VLCDR2 9D9 SAITLAD (SEQ ID NO: 283)VLCDR3 9D9 QHFWNTPYT (SEQ ID NO: 301) VLCDR1 6C1KASQNVGAAVA (SEQ ID NO: 259) VLCDR2 6C1 SASNRYI (SEQ ID NO: 281)VLCDR3 6C1 QQYSNYPLT (SEQ ID NO: 298) VLCDR1 11A10KASQDIHSYLS (SEQ ID NO: 260) VLCDR2 11A10 RTNRLVD (SEQ ID NO: 284)VLCDR3 11A10 LQYDEFPYT (SEQ ID NO: 294) VLCDR1 4A10KASQNVGSAVV (SEQ ID NO: 254) VLCDR2 4A10 SASNRYT (SEQ ID NO: 2)VLCDR3 4A10 QQYSSYPLT (SEQ ID NO: 299) VLCDR1 6A9KASQDINSYLS (SEQ ID NO: 261) VLCDR2 6A9 RANRLVD (SEQ ID NO: 285)VLCDR3 6A9 LQYDEFPYT (SEQ ID NO: 294) VLCDR1 5A2RSSENIYSSLA (SEQ ID NO: 262) VLCDR2 5A2 AATNLAK (SEQ ID NO: 286)VLCDR3 5A2 QHFWGSPFA (SEQ ID NO: 302) VLCDR1 5A5KASQNVGSAVA (SEQ ID NO: 263) VLCDR2 5A5 STSNRYT (SEQ ID NO: 6)VLCDR3 5A5 QQYASYPLT (SEQ ID NO: 303) VLCDR1 12A11RATKGVSKSGYSYMH (SEQ ID NO: 264) VLCDR2 12A11 LASNLES (SEQ ID NO: 276)VLCDR3 12A11 QHSRELPLT (SEQ ID NO: 304) VLCDR1 6D9RASESVDSYGNSLMH (SEQ ID NO: 265) VLCDR2 6D9 IASNLES (SEQ ID NO: 287)VLCDR3 6D9 QQNSEDPRT (SEQ ID NO: 305) VLCDR1 5A3RASESVDRYGNSLMH (SEQ ID NO: 266) VLCDR2 5A3 IASNLES (SEQ ID NO: 287)VLCDR3 5A3 QQNNEDPRT (SEQ ID NO: 306) VLCDR1 6B3KASQDINRYLS (SEQ ID NO: 267) VLCDR2 6B3 RANRLVD (SEQ ID NO: 285)VLCDR3 6B3 LQYDEFPYT (SEQ ID NO: 294) VLCDR1 2C12RASESVDNYGNSFMH (SEQ ID NO: 268) VLCDR2 2C12 LASNLES (SEQ ID NO: 276)VLCDR3 2C12 QQNNEDPRT (SEQ ID NO: 306) VLCDR1 H6A9RATKGVTKSGYSYIH (SEQ ID NO: 269) VLCDR2 H6A9 LASNLQS (SEQ ID NO: 288)VLCDR3 H6A9 QHSRELPLT (SEQ ID NO: 304) VLCDR1 9D12RASKSVDSYGTSFMH (SEQ ID NO: 270) VLCDR2 9D12 LASNLES (SEQ ID NO: 276)VLCDR3 9D12 QQNNEDPRT (SEQ ID NO: 306) VLCDR1 6F2RTSESVDSYGNSFMF (SEQ ID NO: 271) VLCDR2 6F2 LTSNLES (SEQ ID NO: 289)VLCDR3 6F2 QQSNEDPRT (SEQ ID NO: 307) VLCDR1 1A10RASESVDSYGNSFMF (SEQ ID NO: 272) VLCDR2 1A10 LTSNLES (SEQ ID NO: 289)VLCDR3 1A10 QQSNEDPRT (SEQ ID NO: 307) VLCDR1 2A7RASESVDRYGNSLMH (SEQ ID NO: 266) VLCDR2 2A7 IASNLES (SEQ ID NO: 287)VLCDR3 2A7 QQNNEDPRT (SEQ ID NO: 306) VLCDR1 11B7_aRASKSVDSYGTSFMH (SEQ ID NO: 270) VLCDR2 11B7_a LASNLES (SEQ ID NO: 276)VLCDR3 11B7_a QQNNEDPRT (SEQ ID NO: 306) VLCDR1 11B7_bRASKSVDSYGTSFMH (SEQ ID NO: 270) VLCDR2 11B7_b LASNLES (SEQ ID NO: 276)VLCDR3 11B7_b QQNNEDPRT (SEQ ID NO: 306) VLCDR1 7A6RVSESVDRYADSFMH (SEQ ID NO: 273) VLCDR2 7A6 LASNLES (SEQ ID NO: 276)VLCDR3 7A6 QQNKEDPYT (SEQ ID NO: 308) VLCDR1 4D6RASESVDSYGNSFIH (SEQ ID NO: 274) VLCDR2 4D6 RASNLNS (SEQ ID NO: 290)VLCDR3 4D6 HQNNEDPRT (SEQ ID NO: 309) VLCDR1 H2A7RASESIDNYGLIFMS (SEQ ID NO: 275) VLCDR2 H2A7 AASNRGS (SEQ ID NO: 291)VLCDR3 H2A7 QQSKEVPWT (SEQ ID NO: 310) VLCDR1 7F8RASGNIHNYLA (SEQ ID NO: 258) VLCDR2 7F8 NAKTLAD (SEQ ID NO: 292)VLCDR3 7F8 QHFWSTPYT (SEQ ID NO: 311)

TABLE 4 VH CDR Sequences Designation Sequence VHCDR1 9A10GYTFSIFCIH (SEQ ID NO: 18) VHCDR2 9A10 RIDPSSGGTKYNEKFES (SEQ ID NO: 19)VHCDR3 9A10 GEDLLVRTDAMDY (SEQ ID NO: 20) VHCDR1 5A7GYTFSIFCVH (SEQ ID NO: 21) VHCDR2 5A7 RIDPSSGGTKYNEKFES (SEQ ID NO: 19)VHCDR3 5A7 GEDLLVRTDALDY (SEQ ID NO: 22) VHCDR1 5A11GYTFSIFCIH (SEQ ID NO: 23) VHCDR2 5A11 RIDPSSGGTKYNERFEN (SEQ ID NO: 24)VHCDR3 5A11 GEDLLVRTDAMDY (SEQ ID NO: 20) VHCDR1 3B11GYSFSTFFIH (SEQ ID NO: 25) VHCDR2 3B11 RIDPNSGGTKYNEKFES (SEQ ID NO: 26)VHCDR3 3B11 GEDLLIRTDAMDY (SEQ ID NO: 27) VHCDR1 5B9GYSFSTFFIH (SEQ ID NO: 28) VHCDR2 5B9 RIDPNSGATKYNEKFES (SEQ ID NO: 29)VHCDR3 5B9 GEDLLIRTDALDY (SEQ ID NO: 30) VHCDR1 12A7GYTFSTFLIH (SEQ ID NO: 31) VHCDR2 12A7 RIDPNSGGTKYNEKFER (SEQ ID NO: 32)VHCDR3 12A7 GEDLLLRTDAMDY (SEQ ID NO: 33) VHCDR1 9B8GYTFITYWMH (SEQ ID NO: 34) VHCDR2 9B8 GIDPNSGVIKYNEKFKS (SEQ ID NO: 35)VHCDR3 9B8 GEDLLIRTDAMDY (SEQ ID NO: 27) VHCDR1 4B7GYSFSTFCIH (SEQ ID NO: 36) VHCDR2 4B7 RIDPNSGGTKYNEKFES (SEQ ID NO: 26)VHCDR3 4B7 GEDLLIRTDAMDY (SEQ ID NO: 27) VHCDR1 1A4DHIFSIHWMQ (SEQ ID NO: 37) VHCDR2 1A4 EIFPGSGTTDYNEKFKG (SEQ ID NO: 38)VHCDR3 1A4 GAFDY (SEQ ID NO: 39) VHCDR1 5B9ASASGYSFSTFFIH (SEQ ID NO: 81) VHCDR2 5B9ASRIDPNSGATKYNEKFES (SEQ ID NO: 29) VHCDR3 5B9ASGEDLLIRTDALDY (SEQ ID NO: 30) VHCDR1 c04 DHIFSIHWMQ (SEQ ID NO: 37)VHCDR2 c04 EIFPGSGTTDYNEKFKG (SEQ ID NO: 38) VHCDR3 c04GAFDY (SEQ ID NO: 39) VHCDR1 c68 DHIFSIHWMQ (SEQ ID NO: 37) VHCDR2 c68EIFPGSGTTDYNEKFKG (SEQ ID NO: 38) VHCDR3 c68 GAFDY (SEQ ID NO: 39)VHCDR1 c44 DHIFSIHWMQ (SEQ ID NO: 37) VHCDR2 c44EIFPGSGTTDYNEKFKG (SEQ ID NO: 38) VHCDR3 c44 GAFDY (SEQ ID NO: 39)VHCDR1 c03 DHIFSIHWMQ (SEQ ID NO: 37) VHCDR2 c03EIFPGSGTTDYNEKFKG (SEQ ID NO: 38) VHCDR3 c03 GAFDY (SEQ ID NO: 39)VHCDR1 c10 DHIFSIHWMQ (SEQ ID NO: 37) VHCDR2 c10EIFPGSGTTDYNEKFKG (SEQ ID NO: 38) VHCDR3 c10 GAFDY (SEQ ID NO: 39)VHCDR1 c10.1 GYTFTSDDIN (SEQ ID NO: 312) VHCDR2 c10.1WIYPRDGRTKYNEKFKG (SEQ ID NO: 337) VHCDR3 c10.1SRRVYAMDY (SEQ ID NO: 368) VHCDR1 c10.2 GYTFTSDDIN (SEQ ID NO: 312)VHCDR2 c10.2 WIYPRDGRTKYNEKFKG (SEQ ID NO: 337) VHCDR3 c10.2SRRVYAMDY (SEQ ID NO: 368) VHCDR1 8A6 GYTITSYVMH (SEQ ID NO: 313)VHCDR2 8A6 YINPNNDGTKYNEKFKG (SEQ ID NO: 338) VHCDR3 8A6GDYSNYFYWYFDV (SEQ ID NO: 369) VHCDR1 6B7 GYTFPGYWMH (SEQ ID NO: 314)VHCDR2 6B7 KIDPSDSETHYNQNFKD (SEQ ID NO: 339) VHCDR3 6B7EGWDSLTKVWFGW (SEQ ID NO: 370) VHCDR1 4E6 GFNIKDDYMH (SEQ ID NO: 315)VHCDR2 4E6 RIDPAYGNGKYVPKFQD (SEQ ID NO: 340) VHCDR3 4E6RYYAVSSVDYALDY (SEQ ID NO: 371) VHCDR1 3C7 GYIFTSHWMQ (SEQ ID NO: 316)VHCDR2 3C7 DIFPGSGTTDYNEKFKD (SEQ ID NO: 341) VHCDR3 3C7GAFDY (SEQ ID NO: 39) VHCDR1 2A6 GFSLRTFGMGVG (SEQ ID NO: 317)VHCDR2 2A6 HIWWNGDKYYDPALKS (SEQ ID NO: 342) VHCDR3 2A6IGPSITTVAEGFAY (SEQ ID NO: 372) VHCDR1 8A12GFSLRTFGMGVG (SEQ ID NO: 317) VHCDR2 8A12HIWWNDEKYYNPDLKS (SEQ ID NO: 343) VHCDR3 8A12VGPSISTVAEGFPY (SEQ ID NO: 373) VHCDR1 9C5 GFSLRTFGMGVG (SEQ ID NO: 317)VHCDR2 9C5 HIWWNDDKSSHPALKS (SEQ ID NO: 344) VHCDR3 9C5IGPSITTVAEGFAY (SEQ ID NO: 372) VHCDR1 9D1 GFSMRTFGMGVG (SEQ ID NO: 318)VHCDR2 9D1 HIWWNDEKYYNPDLKS (SEQ ID NO: 343) VHCDR3 9D1VGPSISTIAEGFPY (SEQ ID NO: 374) VHCDR1 1B12GYTFTNDNYWMN (SEQ ID NO: 319) VHCDR2 1B12RIRPSDSETHYNQKFTN (SEQ ID NO: 345) VHCDR3 1B12SWEDLLLRSMEDYFDY (SEQ ID NO: 375) VHCDR1 9D9 GFSFSDYNIN (SEQ ID NO: 320)VHCDR2 9D9 KVHPKDGTATYNQKFQD (SEQ ID NO: 346) VHCDR3 9D9LYYDSLTKILFAY (SEQ ID NO: 376) VHCDR1 6C1 GFSLRTFGMGVG (SEQ ID NO: 317)VHCDR2 6C1 HIWWNDEKYYNPALKS (SEQ ID NO: 347) VHCDR3 6C1LGPSITTVAEGFPY (SEQ ID NO: 377) VHCDR1 11A10 GYSLISYYIH (SEQ ID NO: 321)VHCDR2 11A10 LTFPGSGNSKFIEKFKG (SEQ ID NO: 348) VHCDR3 11A10GDFGNYLAYWYFDV (SEQ ID NO: 378) VHCDR1 4A10GFSLKTFGMGVG (SEQ ID NO: 322) VHCDR2 4A10HIWWNDDKFYHPALKS (SEQ ID NO: 349) VHCDR3 4A10IGPSITTVAEGFAY (SEQ ID NO: 372) VHCDR1 6A9 GYSLTSYYIH (SEQ ID NO: 323)VHCDR2 6A9 LIFPGSGNSKYIEKFKG (SEQ ID NO: 350) VHCDR3 6A9GDFGNYLAYWYFDV (SEQ ID NO: 378) VHCDR1 5A2 GFSLNTYGMGVG (SEQ ID NO: 324)VHCDR2 5A2 NIWWNDDKYYNSALKS (SEQ ID NO: 351) VHCDR3 5A2VAATIVTTYGAWFAY (SEQ ID NO: 379) VHCDR1 5A5GFSLRTFGMGVG (SEQ ID NO: 317) VHCDR2 5A5HIWWNDEKYYNPTLKS (SEQ ID NO: 352) VHCDR3 5A5IGPSITTVVEGFPY (SEQ ID NO: 380) VHCDR1 12A11 GYTFTDKYIN (SEQ ID NO: 325)VHCDR2 12A11 WIYPGSGNTKYNEKFKG (SEQ ID NO: 353) VHCDR3 12A11GIIYYYDGSYPYALDY (SEQ ID NO: 381) VHCDR1 6D9 GYTFTDYSIH (SEQ ID NO: 326)VHCDR2 6D9 WIYPGSGNTKYNDKFKG (SEQ ID NO: 354) VHCDR3 6D9DYRRYYAIDY (SEQ ID NO: 382) VHCDR1 5A3 GYTFTDYSIH (SEQ ID NO: 326)VHCDR2 5A3 WIYPGSDNTKYNDKFKG (SEQ ID NO: 355) VHCDR3 5A3DYRRYYAMDY (SEQ ID NO: 383) VHCDR1 6B3 GYTFTSYVMH (SEQ ID NO: 327)VHCDR2 6B3 YLNPNNDGTKYNEKFKG (SEQ ID NO: 356) VHCDR3 6B3GDYSNYFYWYFDV (SEQ ID NO: 369) VHCDR1 2C12 GYTFSDYTIH (SEQ ID NO: 328)VHCDR2 2C12 WIYPGRGNTKYNDKFKG (SEQ ID NO: 357) VHCDR3 2C12DYRRYYAMDY (SEQ ID NO: 383) VHCDR1 H6A9 GYTFTDNFIN (SEQ ID NO: 329)VHCDR2 H6A9 WISPGSGNTKNNEKFKG (SEQ ID NO: 358) VHCDR3 H6A9GIIYYYDGTYPYALDY (SEQ ID NO: 384) VHCDR1 9D12GFSLTDYEIN (SEQ ID NO: 330) VHCDR2 9D12VIWTGGGTKYNSVLIS (SEQ ID NO: 359) VHCDR3 9D12EGRRYYAMDY (SEQ ID NO: 385) VHCDR1 6F2 GFSLTTYEIN (SEQ ID NO: 331)VHCDR2 6F2 VIWTGGTTKYNSAFIS (SEQ ID NO: 360) VHCDR3 6F2EGRRYYAMDY (SEQ ID NO: 385) VHCDR1 11A12 GYTFTSYVIH (SEQ ID NO: 332)VHCDR2 11A12 YLHRNNDGTKYNEKFKV (SEQ ID NO: 361) VHCDR3 11A12GDYSNYFYWYFDV (SEQ ID NO: 386) VHCDR1 1A10 GFSLTTYEIN (SEQ ID NO: 331)VHCDR2 1A10 VIWTGGTTKYNSAFIS (SEQ ID NO: 362) VHCDR3 1A10EGRRYYAMDY (SEQ ID NO: 385) VHCDR1 2A7 GYTFTDYSIH (SEQ ID NO: 326)VHCDR2 2A7 WIYPGSDNTKYNDKFKG (SEQ ID NO: 355) VHCDR3 2A7DYRRYYAMDY (SEQ ID NO: 383) VHCDR1 11B7_a GFSLTDYEIN (SEQ ID NO: 330)VHCDR2 11B7_a VIWTGGGTKYNSVLIS (SEQ ID NO: 359) VHCDR3 11B7_aEGRRYYAMDY (SEQ ID NO: 385) VHCDR1 11B7_b GFSLSTFGMGVG (SEQ ID NO: 392)VHCDR2 11B7_b HIWWDDDKYYNPALKS (SEQ ID NO: 363) VHCDR3 11B7_bIEGPYYWYFDV (SEQ ID NO: 387) VHCDR1 7A6 GFSLSRYSVH (SEQ ID NO: 333)VHCDR2 7A6 MIWGGGNTDYNSGLKS (SEQ ID NO: 364) VHCDR3 7A6PSLYYYDVAWFPY (SEQ ID NO: 388) VHCDR1 4D6 GFNIKNDYLH (SEQ ID NO: 334)VHCDR2 4D6 WIDSANDKTKYAPKFQD (SEQ ID NO: 365) VHCDR3 4D6VGVQDGYYVRDFDY (SEQ ID NO: 389) VHCDR1 H2A7 GFTFSDYYMS (SEQ ID NO: 335)VHCDR2 H2A7 LIRNKAPGYTTEYSASVKG  (SEQ ID NO: 366) VHCDR3 H2A7VLRRADCLDY (SEQ ID NO: 390) VHCDR1 H7F8 GFTFSSYDMS (SEQ ID NO: 336)VHCDR2 H7F8 AINSYGVNTYYPDTVKD (SEQ ID NO: 367) VHCDR3 H7F8LLIGPYYYAMDY (SEQ ID NO: 391)

The CDRs listed above in Tables 3 and 4 are defined using the Chothianumbering system (Al-Lazikani et al., JMB, 273, 927-948, (1997)) or IGMTnumbering system (Lefranc, M.-P. et al., Dev. Comp. Immunol., 27, 55-77(2003)). The IgAligner IMGT algorithm is from Chemical Computing Group(CCG).

Fabs that showed better or equal binding as compared to the chimericparent Fab were selected for conversion to IgG. was converted to anIgG1KO format. IgG1KO (knock-out of effector functions) has twomutations in the Fc region, Leu234Ala and Leu235Ala, which reduceeffector function such as FcγR and complement binding. The IgG format isdescribed in the literature (see for example Hezareh et al. (2001)Journal of Virology 75: 12161-12168). Example 2 describes thehumanization process in further detail. The results of such humanizationresulted in humanized antibody sequences. A representative number ofhumanized light chain and heavy chain variable regions derived frommouse antibodies 5B9 and 1A4 are provided and shown in Tables 5 and 6.

TABLE 5 Humanized 5B9 and 1A4 Vκ Sequences Designation Sequence 148c04VKDIQMTQSPSSLSASVGDRVTITCRASQDIGNRLSWLQQEPGKAPKRLIYATSSLDSGVPSRFSGSRSGTEFTLTISSLQPEDFVTYYCLQYASSPFTFGQGTKLEIK (SEQ ID NO: 82) GACATCCAGATGACCCAGAGCCCAAGCAGCCTGAGCGCCAGCGTCGGCGACCGCGTGACCATCACCTGCCGCGCCAGCCAGGACATCGGCAACCGCCTGTCGTGGCTGCAGCAGGAGCCAGGCAAGGCCCCAAAGCGCCTGATCTACGCCACCAGCAGCCTGGACAGCGGTGTCCCAAGCCGCTTCAGCGGCAGCCGCAGCGGCACCGAGTTCACCCTGACCATCAGCAGCCTGCAACCAGAGGACTTCGTCACCTACTACTGCCTGCAATACGCCAGCAGCCCATTCACCTTCGGCCAGGGCACCAAGCTGGAGATCAAG (SEQ ID NO: 234) 148c18VKDIQMTQSPSSLSASVGDRVTITCRASQDIGNRLNWLQQEPGKAPKRLIYATSSLDSGVPKRFSSSRSGTEFTLTISSLQPEDFVDYYCLQYASSPFTFGTGTKLEIK (SEQ ID NO: 83) 148c19VKDIQMTQSPSSLSASVGDRVTITCRASQDIGNRLNWYQQEPGKAPKRLIYATSSLDSGVPKRFSGSRSGTEFTLTISSLQPEDFVDYYCLQYASSPFTFGTGTKLEIK (SEQ ID NO: 84) 148c68VKDIQMTQSPSSLSASVGDRVTITCRASQDIGNRLHWYQQKPGKAPKRLIYATSSLDSGVPKRFSGSRSGTEFTLTISSLQPEDFVTYYCLQYASSPFTFGQGTKLEIK (SEQ ID NO: 85) 148c77VKDIQMTQSPSSLSASVGDRVTITCRASQDIGNRLNWYQQKPGKAPKRLIYATSSLDSGVPKRFSGSRSGTEFTLTISSLQPEDFVDYYCLQYASSPFTFGTGTKLEIK (SEQ ID NO: 86) 148c92VKDIQMTQSPSSLSASVGDRVTITCRASQDIGNRLNWYQQKPGKAPKRLIYATSSLDSGVPSRFSGSRSGTEFTLTISSLQPEDFVDYYCLQYASSPFTFGTGTKLEIK (SEQ ID NO: 87) 160c16VKDIQMTQSPSSLSASVGDRVTITCRASQDIGNRLNWYQQKPGKAPKRLIYATSSLDSGVPSRFSGSRSGTEFTLTISSLQPEDFVTYYCLQYASSPFTFGQGTKLEIK (SEQ ID NO: 88) GACATCCAGATGACCCAGAGCCCAAGCAGCCTGAGCGCCAGCGTCGGCGACCGCGTGACCATCACCTGCCGCGCCAGCCAGGACATCGGCAACCGCCTGAACTGGTACCAGCAGAAGCCAGGCAAGGCCCCAAAGCGCCTGATCTACGCCACCAGCAGCCTGGACAGCGGTGTCCCAAGCCGCTTCAGCGGCAGCCGCAGCGGCACCGAGTTCACCCTGACCATCAGCAGCCTGCAACCAGAGGACTTCGTCACCTACTACTGCCTGCAATACGCCAGCAGCCCATTCACCTTCGGCCAGGGCACCAAGCTGGAGATCAAG (SEQ ID NO: 393) 148c44VKDIQMTQSPSSLSASVGDRVTITCRASQDIGNRLPWLQQKPGKAPKRLIYATSSLDSGVPSRFSGSGSGTEFTLTISSLQPEDFVDYYCLQYASSPFTFGTGTKLEIK (SEQ ID NO: 89) 148c03VKDIQMTQSPSSLSASVGDRVTITCRASQDIGNRLRWYQQKPGKAPKRLIYATSSLDSGVPSRFSGSRSGTEFTLTISSLQPEDFATYYCLQYASSPFTFGQGTKLEIK (SEQ ID NO: 90) 148c10VKDIQMTQSPSSLSASVGDRVTITCRASQDIGNRLMWYQQKPGKAPKRLIYATSSLDSGVPSRFSGSRSGTEFTLTISSLQPEDFVTYYCLQYASSPFTFGTGTKLEIK (SEQ ID NO: 91) 145c02VKDIQMTQSPSSLSASVGDRVTITCKASQNAGIDVAWFQQKPGKAPKLLIYSKSNRYTGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCLQYRSYPRTFGQGTKLEIK (SEQ ID NO: 92) 145c08VKDIQMTQSPSSLSASVGDRVSITCKASQNAGIDVAWFQQKPGKAPKLLIYSKSNRYTGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCLQYRSYPRTFGQGTKLEIK (SEQ ID NO: 93) GACATCCAGATGACCCAGAGCCCAAGCAGCCTGAGCGCCAGCGTCGGCGACCGCGTGAGCATCACCTGCAAGGCCAGCCAGAACGCCGGCATCGACGTGGCTTGGTTCCAGCAGAAGCCTGGCAAGGCCCCAAAGCTGCTGATCTACAGCAAGAGCAACCGCTACACGGCGTGCCAAGCCGCTTCAGCGGCAGCGGCAGCGGCACCGACTTCACCCTGACCATCAGCAGCCTCCAGCCAGAGGACTTCGCCACCTACTACTGCCTCCAGTACCGCAGCTACCCACGCACCTTCGGCCAGGGCACCAAGCTGGAGATCAAG (SEQ ID NO: 394) 145c15VKDIQMTQSPSSLSASVGDRVSITCKASQNAGIDVAWFQQKPGKAPKLLIYSKSNRYTGVPSRFSGSGSGTDFTLTISSLQPEDFADYYCLQYRSYPRTFGGGTKLEIK (SEQ ID NO: 94) GACATCCAGATGACCCAGAGCCCAAGCAGCCTGAGCGCCAGCGTCGGCGACCGCGTGAGCATCACCTGCAAGGCCAGCCAGAACGCCGGCATCGACGTGGCTTGGTTCCAGCAGAAGCTGGCAAGGCCCCAAAGCTGCTGATCTACAGCAAGAGCAACCGCTACACCGGCGTGCCAAGCCGCTTCAGCGGCAGCGGCAGCGGCACCGACTTCACCCTGACCATCAGCAGCCTCCAGCCAGAGGACTTCGCCGACTACTACTGCCTCCAGTACCGCAGCTACCCACGCACCTTCGGCGGCGGCACCAAGCTGGAGATCAAG (SEQ ID NO: 395) 145c18VKDIQMTQSPSSLSASVGDRVSITCKASQNAGIDVAWFQQKPGKAPKLLIFSKSNRYTGVPDRFSGSGSGTDFTLTISSLQPEDFADYYCLQYRSYPRTFGQGTKLEIK (SEQ ID NO: 95) 145c28VKDIQMTQSPSSLSASVGDRVTITCKASQNAGIDVAWFQQKPGKAPKLLIFSKSNRYTGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCLQYRSYPRTFGQGTKLEIK (SEQ ID NO: 96) 145c36VKDIVMTQSPSSLSASVGDRVSITCKASQNAGIDVAWFQQKPGKAPKLLIYSKSNRYTGVPDRFSGSGSGTDFTLTISSLQPEDFATYYCLQYRSYPRTFGQGTKLEIK (SEQ ID NO: 97) Reference 1EIVLTQSPATLSLSPGERATLSCRASQSVSRYLAWYQQKPGQAPRLLIYDASNRATGIPARFSGSGSGTDSTLTISSLEPEDFAVYYCQQRSNWPRTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO: 98) Reference 2SSELTQDPAVSVALGQTVRVTCQGDSLRSYYASWYQQKPGQAPVLVIYGKNNRPSGIPDRFSGSSSGNTASLTITGAQAEDEADYYCSSRDSSGNHWVFGGGTELTVLGQPKAAPSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNNKYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS (SEQ ID NO: 99)

TABLE 6 Humanized 5B9 and 1A4 VH Sequences Designation Sequence 148c04VHQVQLVQSGAEVKKPGSSVKVSCKAPDHIFSIHWMQWVRQRPGQGLEWIGEIFPGSGTTDYNEKFKGKVTITVDKSTSTAYMELSSLRSEDTAVYFCASGAFDYWGQGTTVTVSS (SEQ ID NO: 100) 148c18VHQVQLVQSGAEVKKPGSSVKVSCKAPDHIFSIHWMQWVRQAPGQGLEWMGEIFPGSGTTDYNEKFKGKVTITVDKSTSTAYMELSSLRSEDTAVYYCASGAFDYWGQGTTVTVSS (SEQ ID NO: 101)CAGGTGCAGCTGGTGCAGAGCGGCGCCGAGGTGAAGAAGCCAGGCAGCAGCGTGAAGGTCAGCTGCAAGGCCCCCGACCACATCTTCAGCATCCACTGGATGCAGTGGGTCCGCCAAGCCCCAGGCCAGGGCCTGGAGTGGATGGGCGAGATTTTCCCAGGCAGCGGCACCACCGACTACAACGAGAAGTTCAAGGGCAAGGTGACCATCACCGTCGACAAGAGCACCAGCACCGCCTACATGGAGCTGAGCAGCCTGCGCAGCGAGGACACCGCCGTCTACTACTGCGCCAGCGGCGCCTTCGACTACTGGGGCCAGGGCACCACCGTGACCGTGAGCAGC (SEQ ID NO: 396) 148c19VHQVQLVQSGAEVKKPGSSVKISCKAPDHIFSIHWMQWVRQRPGQGLEWIGEIFPGSGTTDYNEKFKGKVTVTVDKSTSTAYMELSSLRSEDTAVYYCASGAFDYWGQGTTVTVSS (SEQ ID NO: 102) 148c68VHQVQLVQSGAEVKKPGSSVKISCKASDHIFSIHWMQWVRQRPGQGLEWIGEIFPGSGTTDYNEKFKGKVTVTVDKSTSTAYMELSSLRSEDTAVYFCARGAFDYWGQGTTVTVSS (SEQ ID NO: 103) 148c75VHQVQLVQSGAEVKKPGSSVKVSCKAPDHIFSIHWMQWVRQRPGQGLEWMGEIFPGSGTTDYNEKFKGKVTVTADKSTSTAYMELSSLRSEDTAVYFCASGAFDYWGQGTTVTVSS (SEQ ID NO: 104) 148c77VHQVQLVQSGAEVKKPGSSVKISCKASDHIFSIHWMQWVRQAPGQGLEWMGEIFPGSGTTDYNEKFKGKVTVTVDKSTSTAYMELSSLRSEDTAVYFCASGAFDYWGQGTTVTVSS (SEQ ID NO: 105) 148c92VHQVQLVQSGAEVKKPGSSVKVSCKAPDHIFSIHWMQWVRQAPGQGLEWIGEIFPGSGTTDYNEKFKGRATVTVDKSTSTAYMELSSLRSEDTAVYFCASGAFDYWGQGTTVTVSS (SEQ ID NO: 106) 161c01VHQVQLVQSGAEVKKPGSSVKVSCKASDHIFSIHWMQWVRQAPGQGLEWMGEIFPGSGTTDYNEKFKGRVTITADKSTSTAYMELSSLRSEDTAVYYCARGAFDYWGQGTTVTVSS (SEQ ID NO: 107) 148c03VHQVQLVQSGAEVKKPGSSVKISCKAPDHIFSIHWMQWVRQAPGQGLEWIGEIFPGSGTTDYNEKFKGKVTVTVDKSTSTAYMELSSLRSEDTAVYYCASGAFDYWGQGTTVTVSS (SEQ ID NO: 108) 148c10VHQVQLVQSGAEVKKPGSSVKISCKASDHIFSIHWMQWVRQRPGQGLEWIGEIFPGSGTTDYNEKFKGKVTITADKSTSTAYMELSSLRSEDTAVYYCASGAFDYWGQGTTVTVSS (SEQ ID NO: 109) 145c02VHQVQLVQSGAEVKKPGASVKVSCKASGYSFSTFFIHWIQQRPGQGLEWMGRIDPNSGATKYNEKFESRVTMTRDTSISTAYMELSRLRSDDTAVYYCARGEDLLIRTDALDYWGQGTLVTVSS (SEQ ID NO: 110) 145c08VHQVQLVQSGAEVKKPGASVKVSCKASGYSFSTFFIHWIQQRPGQGLEWMGRIDPNSGATKYNEKFESKVTLTVDTSISTAYMELSRLRSDDTAVYYCARGEDLLIRTDALDYWGQGTSVTVSS (SEQ ID NO: 111) 145c15VHQVQLVQSGAEVKKPGASVKVSCKASGYSFSTFFIHWVRQRPGQGLEWIGRIDPNSGATKYNEKFESKVTLTRDTSISTAYMELSRLRSDDTAVYYCARGEDLLIRTDALDYWGQGTSVTVSS (SEQ ID NO: 112)CAGGTGCAGCTGGTGCAGAGCGGCGCTGAGGTGAAGAAGCCAGGCGCCAGCGTGAAGGTGAGCTGCAAGGCCAGCGGCTACAGCTTCAGCACCTTCTTCATCCACTGGGTCCGCCAACGCCCAGGCCAGGGCCTGGAGTGGATCGGCCGCATCGACCCAAACAGCGGCGCCACCAAGTACAACGAGAAGTTCGAGAGCAAGGTCACCCTGACCCGCGACACCAGCATCAGCACCGCCTACATGGAGCTGAGCCGCCTGCGCAGCGACGACACCGCCGTCTACTACTGCGCCCGCGGCGAGGACCTGCTGATCCGCACCGACGCCCTGGATTACTGGGGTCAGGGTACTAGCGTGACCGTGAGCAGC (SEQ ID NO: 397) 145c18VHQVQLVQSGAEVKKPGASVKVSCKASGYSFSTFFIHWVQQRPGQGLEWMGRIDPNSGATKYNEKFESKVTLTRDTSISTAYMELSRLRSDDTAVYYCARGEDLLIRTDALDYWGQGTLVTVSS (SEQ ID NO: 113) 145c28VHQVQLVQSGAEVKKPGASVKVSCKASGYSFSTFFIHWVRQAPGQGLEWIGRIDPNSGATKYNEKFESRVTMTRDTSISTAYMELSRLRSDDTAVYYCARGEDLLIRTDALDYWGQGTSVTVSS (SEQ ID NO: 114)CAGGTGCAGCTGGTGCAGAGCGGCGCTGAGGTGAAGAAGCCAGGCGCCAGCGTGAAGGTGAGCTGCAAGGCCAGCGGCTACAGCTTCAGCACCTTCTTCATCCACTGGGTCCGCCAAGCCCCAGGCCAGGGCCTGGAGTGGATCGGCCGCATCGACCCAAACAGCGGCGCCACCAAGTACAACGAGAAGTTCGAGAGCCGCGTCACCATGACCCGCGACACCAGCATCAGCACCGCCTACATGGAGCTGAGCCGCCTGCGCAGCGACGACACCGCCGTCTACTACTGCGCCCGCGGCGAGGACCTGCTGATCCGCACCGACGCCCTGGATTACTGGGGTCAGGGTACTAGCGTGACCGTGAGCAGC (SEQ ID NO: 398) 145c36VHQVQLVQSGAEVKKPGASVKVSCKASGYSFSTFFIHWVRQRPGQGLEWMGRIDPNSGATKYNEKFESRATLTVDTSISTAYMELSRLRSDDTAVYYCARGEDLLIRTDALDYWGQGTSVTVSS (SEQ ID NO: 115) Reference 1QVQLQQWGAGLLKPSETLSLTCAVYGGSFSGYYWSWIRQPPGKGLEWIGEINHSGSTNYNPSLKSRVTISVDTSKNQFSLKLSSVTAADTAVYYCARGYYDILTGYYYYFDYWGQGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYGPPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKTISKAKGQPREPQVYTLPPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLG K (SEQ ID NO: 116)Reference 2 QVQLQQSGAEVKKPGSSVRVSCKASGGTFNNNAINWVRQAPGQGLEWMGGIIPMFGTAKYSQNFQGRVAITADESTGTASMELSSLRSEDTAVYYCARSRDLLLFPHHALSPWGRGTMVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLS LSPG (SEQ ID NO: 117)

The humanized anti-BAFF antibodies optionally include specific aminoacid substitutions in the consensus or germline framework regions. Thespecific substitution of amino acid residues in these frameworkpositions can improve various aspects of antibody performance includingbinding affinity and/or stability, over that demonstrated in humanizedantibodies formed by “direct swap” of CDRs or HVLs into the humangermline framework regions.

In some embodiments, the present invention describes other monoclonalantibodies with a light chain variable region having the amino acidsequence set forth in of SEQ ID NO: 41, 43, 45, 47, 49, 51, 53, 55, 57,119, 121, 123, 125, 127, 129, 131, 133, 135, 137, 139, 141, 143, 145,147, 149, 151, 153, 155, 157, 159, 161, 163, 165, 167, 169, 171, 173,175, 177, 179 or 181. In some embodiments, the present inventiondescribes other monoclonal antibodies with a heavy chain variable regionhaving the amino acid sequence set forth in of SEQ ID NO: 59, 61, 63,65, 67, 69, 71, 73, 75, 183, 185, 187, 189, 191, 193, 195, 197, 199,201, 203, 205, 207, 209, 211, 213, 215, 217, 219, 221, 223, 225, 227,229, 231, 233, 235, 238, 240, 242, 244, 246 or 248 (see Tables 1 and 2above). The CDR sequence of these mouse antibodies are shown in Tables 3and 4. Placing such CDRs into FRs of the human consensus heavy and lightchain variable domains will yield useful humanized antibodies of thepresent invention.

In particular, the present invention provides monoclonal antibodies withthe combinations of light chain variable and heavy chain variableregions of SEQ ID NO: 41/59, 43/61, 45/63, 47/65, 49/67, 51/69, 53/71,55/73, 57/75, 119/183, 121/185, 123/187, 125/189, 127/191, 129/193,131/195, 133/197, 135/199, 137/201, 139/203, 141/205, 143/207, 145/209,147/211, 149/213, 151/215, 153/217, 155/219, 157/221, 159/223, 161/225,163/227, 165/229, 167/231, 169/233, 171/235, 173/238, 173/240, 175/242,177/244, 179/246 or 181/248. Such variable regions can be combined withhuman constant regions.

In some embodiments, the present invention describes other humanizedantibodies with light chain variable region sequences having the aminoacid sequence set forth in of SEQ ID NO: 82-97. In some embodiments, thepresent invention describes other humanized antibodies with heavy chainvariable region sequences having the amino acid sequence set forth in ofSEQ ID NO: 100-115 (see Tables 5 and 6 above). The CDR sequences ofthese antibodies are shown in Tables 3 and 4. Such variable regions canbe combined with human constant regions.

In some specific embodiments, the humanized anti-BAFF antibodiesdisclosed herein comprise at least a heavy or a light chain variabledomain comprising the CDRs or HVLs of the murine monoclonal antibodiesor humanized antibodies as shown in Tables 1 through 6 above and the FRsof the human germline heavy and light chain variable domains.

The CDRs of these sequences are shown in Tables 3 and 4. In oneembodiment, the invention provides an anti-BAFF antibody moleculecomprising a light chain variable domain with a CDR1 selected from thegroup consisting of SEQ ID NO: 1, SEQ ID NO: 5, SEQ ID NO: 10, SEQ IDNO: 13, SEQ ID NO: 15, SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78, SEQID NO: 79, SEQ ID NO: 80, SEQ ID NO: 249, SEQ ID NO: 250, SEQ ID NO:251, SEQ ID NO: 252, SEQ ID NO: 253, SEQ ID NO: 254, SEQ ID NO: 255, SEQID NO: 256, SEQ ID NO: 257, SEQ ID NO: 258, SEQ ID NO: 259, SEQ ID NO:260, SEQ ID NO: 261, SEQ ID NO: 262, SEQ ID NO: 263, SEQ ID NO: 264, SEQID NO: 265, SEQ ID NO: 266, SEQ ID NO: 267, SEQ ID NO: 268, SEQ ID NO:269, SEQ ID NO: 270, SEQ ID NO: 271, SEQ ID NO: 272, SEQ ID NO: 273, SEQID NO: 274 and SEQ ID NO: 275; a CDR2 selected from the group consistingof SEQ ID NO: 2, SEQ ID NO: 6, SEQ ID NO: 8, SEQ ID NO: 11, SEQ ID NO:16, SEQ ID NO: 276, SEQ ID NO: 277, SEQ ID NO: 278, SEQ ID NO: 279, SEQID NO: 280, SEQ ID NO: 281, SEQ ID NO: 282, SEQ ID NO: 283, SEQ ID NO:284, SEQ ID NO: 285, SEQ ID NO: 286, SEQ ID NO: 287, SEQ ID NO: 288, SEQID NO: 289, SEQ ID NO: 290, SEQ ID NO: 291 and SEQ ID NO: 292; and aCDR3 selected from the group consisting of SEQ ID NO: 3, SEQ ID NO: 4,SEQ ID NO: 7, SEQ ID NO: 9, SEQ ID NO: 12, SEQ ID NO:14, SEQ ID NO: 17,SEQ ID NO: 293, SEQ ID NO: 294, SEQ ID NO: 295, SEQ ID NO: 296, SEQ IDNO: 297, SEQ ID NO: 298, SEQ ID NO: 299, SEQ ID NO: 300, SEQ ID NO: 301,SEQ ID NO: 302, SEQ ID NO: 303, SEQ ID NO: 304, SEQ ID NO: 305, SEQ IDNO: 306, SEQ ID NO: 307, SEQ ID NO: 308, SEQ ID NO: 309, SEQ ID NO: 310and SEQ ID NO: 311; and a heavy chain variable domain with a CDR1selected from the group consisting of SEQ ID NO: 18, SEQ ID NO: 21, SEQID NO: 23, SEQ ID NO: 25, SEQ ID NO: 28, SEQ ID NO: 31, SEQ ID NO: 34,SEQ ID NO: 36, SEQ ID NO: 37, SEQ ID NO: 81, SEQ ID NO: 312, SEQ ID NO:313, SEQ ID NO: 314, SEQ ID NO: 315, SEQ ID NO: 316, SEQ ID NO: 317, SEQID NO: 318, SEQ ID NO: 319, SEQ ID NO: 320, SEQ ID NO: 321, SEQ ID NO:322, SEQ ID NO: 323, SEQ ID NO: 324, SEQ ID NO: 325, SEQ ID NO: 326, SEQID NO: 327, SEQ ID NO: 328, SEQ ID NO: 329, SEQ ID NO: 330, SEQ ID NO:331, SEQ ID NO: 332, SEQ ID NO: 392, SEQ ID NO: 333, SEQ ID NO: 334, SEQID NO: 335 and SEQ ID NO: 336; a CDR2 selected from the group consistingof SEQ ID NO: 19, SEQ ID NO: 24, SEQ ID NO: 26, SEQ ID NO: 29, SEQ IDNO: 32, SEQ ID NO: 35, SEQ ID NO: 38, SEQ ID NO: 337, SEQ ID NO: 338,SEQ ID NO: 339, SEQ ID NO: 340, SEQ ID NO: 341, SEQ ID NO: 342, SEQ IDNO: 343, SEQ ID NO: 344, SEQ ID NO: 343, SEQ ID NO: 345, SEQ ID NO: 346,SEQ ID NO: 347, SEQ ID NO: 348, SEQ ID NO: 349, SEQ ID NO: 350, SEQ IDNO: 351, SEQ ID NO: 352, SEQ ID NO: 353, SEQ ID NO: 354, SEQ ID NO: 355,SEQ ID NO: 356, SEQ ID NO: 357, SEQ ID NO: 358, SEQ ID NO: 359, SEQ IDNO: 360, SEQ ID NO: 361, SEQ ID NO: 362, SEQ ID NO: 363, SEQ ID NO: 364,SEQ ID NO: 365, SEQ ID NO: 366 and SEQ ID NO: 367; and a CDR3 selectedfrom the group consisting of SEQ ID NO: 20, SEQ ID NO: 22, SEQ ID NO:27, SEQ ID NO: 30, SEQ ID NO: 33, SEQ ID NO. 39, SEQ ID NO: 368, SEQ IDNO: 369, SEQ ID NO: 370, SEQ ID NO: 371, SEQ ID NO: 372, SEQ ID NO: 373,SEQ ID NO: 374, SEQ ID NO: 375, SEQ ID NO: 376, SEQ ID NO: 377, SEQ IDNO: 378, SEQ ID NO: 378, SEQ ID NO: 379, SEQ ID NO:380, SEQ ID NO: 381,SEQ ID NO: 382, SEQ ID NO: 383, SEQ ID NO: 384, SEQ ID NO: 385, SEQ IDNO: 386, SEQ ID NO: 387, SEQ ID NO: 388, SEQ ID NO: 389, SEQ ID NO: 390and SEQ ID NO: 391.

In another embodiment, the invention provides (a) an anti-BAFF antibodymolecule where the light chain variable domain comprises a CDR1 of SEQID NO: 1, a CDR2 of SEQ ID NO: 2 and a CDR3 of SEQ ID NO: 3 and a heavychain variable domain comprises a CDR1 of SEQ ID NO: 18, a CDR2 of SEQID NO: 19 and a CDR3 of SEQ ID NO: 20; (b) an anti-BAFF antibodymolecule where the light chain variable domain comprises a CDR1 of SEQID NO: 1, a CDR2 of SEQ ID NO: 2 and a CDR3 of SEQ ID NO: 4 and a heavychain variable domain comprises a CDR1 of SEQ ID NO: 21, a CDR2 of SEQID NO: 19 and a CDR3 of SEQ ID NO: 22; (c) an anti-BAFF antibodymolecule where the light chain variable domain comprises a CDR1 of SEQID NO: 1, a CDR2 of SEQ ID NO: 2 and a CDR3 of SEQ ID NO: 4 and a heavychain variable domain comprises a CDR1 of SEQ ID NO: 23, a CDR2 of SEQID NO: 24 and a CDR3 of SEQ ID NO: 20; (d) an anti-BAFF antibodymolecule where the light chain variable domain comprises a CDR1 of SEQID NO: 5, a CDR2 of SEQ ID NO: 6 and a CDR3 of SEQ ID NO: 7 and a heavychain variable domain comprises a CDR1 of SEQ ID NO: 25, a CDR2 of SEQID NO: 26 and a CDR3 of SEQ ID NO: 27; (e) an anti-BAFF antibodymolecule where the light chain variable domain comprises a CDR1 of SEQID NO: 5, a CDR2 of SEQ ID NO: 8 and a CDR3 of SEQ ID NO: 9 and a heavychain variable domain comprises a CDR1 of SEQ ID NO: 28, a CDR2 of SEQID NO: 29 and a CDR3 of SEQ ID NO: 30; (f) an anti-BAFF antibodymolecule where the light chain variable domain comprises a CDR1 of SEQID NO: 10, a CDR2 of SEQ ID NO: 11 and a CDR3 of SEQ ID NO: 12 and aheavy chain variable domain comprises a CDR1 of SEQ ID NO: 31, a CDR2 ofSEQ ID NO: 32 and a CDR3 of SEQ ID NO: 33; (g) an anti-BAFF antibodymolecule where the light chain variable domain comprises a CDR1 of SEQID NO: 13, a CDR2 of SEQ ID NO: 6 and a CDR3 of SEQ ID NO: 14 and aheavy chain variable domain comprises a CDR1 of SEQ ID NO: 34, a CDR2 ofSEQ ID NO: 35 and a CDR3 of SEQ ID NO: 27; (h) an anti-BAFF antibodymolecule where the light chain variable domain comprises a CDR1 of SEQID NO: 10, a CDR2 of SEQ ID NO: 6 and a CDR3 of SEQ ID NO: 7 and a heavychain variable domain comprises a CDR1 of SEQ ID NO: 36, a CDR2 of SEQID NO: 26 and a CDR3 of SEQ ID NO: 27; (i) an anti-BAFF antibodymolecule where the light chain variable domain comprises a CDR1 of SEQID NO: 15, a CDR2 of SEQ ID NO: 16 and a CDR3 of SEQ ID NO: 17 and aheavy chain variable domain comprises a CDR1 of SEQ ID NO: 37, a CDR2 ofSEQ ID NO: 38 and a CDR3 of SEQ ID NO: 39; (j) an anti-BAFF antibodymolecule where the light chain variable domain comprises a CDR1 of SEQID NO: 76, a CDR2 of SEQ ID NO: 16 and a CDR3 of SEQ ID NO: 17 and aheavy chain variable domain comprises a CDR1 of SEQ ID NO: 37, a CDR2 ofSEQ ID NO: 38 and a CDR3 of SEQ ID NO: 39; (k) an anti-BAFF antibodymolecule where the light chain variable domain comprises a CDR1 of SEQID NO: 77, a CDR2 of SEQ ID NO: 16 and a CDR3 of SEQ ID NO: 17 and aheavy chain variable domain comprises a CDR1 of SEQ ID NO: 37, a CDR2 ofSEQ ID NO: 38 and a CDR3 of SEQ ID NO: 39; (l) an anti-BAFF antibodymolecule where the light chain variable domain comprises a CDR1 of SEQID NO: 78, a CDR2 of SEQ ID NO: 16 and a CDR3 of SEQ ID NO: 17 and aheavy chain variable domain comprises a CDR1 of SEQ ID NO: 37, a CDR2 ofSEQ ID NO: 38 and a CDR3 of SEQ ID NO: 39; (m) an anti-BAFF antibodymolecule where the light chain variable domain comprises a CDR1 of SEQID NO: 79, a CDR2 of SEQ ID NO: 16 and a CDR3 of SEQ ID NO: 17 and aheavy chain variable domain comprises a CDR1 of SEQ ID NO: 37, a CDR2 ofSEQ ID NO: 38 and a CDR3 of SEQ ID NO: 39; (n) an anti-BAFF antibodymolecule where the light chain variable domain comprises a CDR1 of SEQID NO: 80, a CDR2 of SEQ ID NO: 16 and a CDR3 of SEQ ID NO: 17 and aheavy chain variable domain comprises a CDR1 of SEQ ID NO: 37, a CDR2 ofSEQ ID NO: 38 and a CDR3 of SEQ ID NO: 39; (o) an anti-BAFF antibodymolecule where the light chain variable domain comprises a CDR1 of SEQID NO: 5, a CDR2 of SEQ ID NO: 8 and a CDR3 of SEQ ID NO: 9 and a heavychain variable domain comprises a CDR1 of SEQ ID NO: 81, a CDR2 of SEQID NO: 29 and a CDR3 of SEQ ID NO: 30; (p) an anti-BAFF antibodymolecule where the light chain variable domain comprises a CDR1 of SEQID NO: 249, a CDR2 of SEQ ID NO: 276 and a CDR3 of SEQ ID NO: 293 and aheavy chain variable domain comprises a CDR1 of SEQ ID NO: 312, a CDR2of SEQ ID NO: 337 and a CDR3 of SEQ ID NO: 368; (q) an anti-BAFFantibody molecule where the light chain variable domain comprises a CDR1of SEQ ID NO: 250, a CDR2 of SEQ ID NO: 276 and a CDR3 of SEQ ID NO: 293and a heavy chain variable domain comprises a CDR1 of SEQ ID NO: 312, aCDR2 of SEQ ID NO: 337 and a CDR3 of SEQ ID NO: 368; (r) an anti-BAFFantibody molecule where the light chain variable domain comprises a CDR1of SEQ ID NO: 251, a CDR2 of SEQ ID NO: 277 and a CDR3 of SEQ ID NO: 294and a heavy chain variable domain comprises a CDR1 of SEQ ID NO: 313, aCDR2 of SEQ ID NO: 338 and a CDR3 of SEQ ID NO: 369; (s) an anti-BAFFantibody molecule where the light chain variable domain comprises a CDR1of SEQ ID NO: 252, a CDR2 of SEQ ID NO: 278 and a CDR3 of SEQ ID NO: 295and a heavy chain variable domain comprises a CDR1 of SEQ ID NO: 314, aCDR2 of SEQ ID NO: 339 and a CDR3 of SEQ ID NO: 370; (t) an anti-BAFFantibody molecule where the light chain variable domain comprises a CDR1of SEQ ID NO: 253, a CDR2 of SEQ ID NO: 279 and a CDR3 of SEQ ID NO: 296and a heavy chain variable domain comprises a CDR1 of SEQ ID NO: 315, aCDR2 of SEQ ID NO: 340 and a CDR3 of SEQ ID NO: 371; (u) an anti-BAFFantibody molecule where the light chain variable domain comprises a CDR1of SEQ ID NO: 15, a CDR2 of SEQ ID NO: 16 and a CDR3 of SEQ ID NO: 297and a heavy chain variable domain comprises a CDR1 of SEQ ID NO: 316, aCDR2 of SEQ ID NO: 341 and a CDR3 of SEQ ID NO: 39; (v) an anti-BAFFantibody molecule where the light chain variable domain comprises a CDR1of SEQ ID NO: 254, a CDR2 of SEQ ID NO: 280 and a CDR3 of SEQ ID NO: 298and a heavy chain variable domain comprises a CDR1 of SEQ ID NO: 317, aCDR2 of SEQ ID NO: 342 and a CDR3 of SEQ ID NO: 372; (w) an anti-BAFFantibody molecule where the light chain variable domain comprises a CDR1of SEQ ID NO: 255, a CDR2 of SEQ ID NO: 281 and a CDR3 of SEQ ID NO: 298and a heavy chain variable domain comprises a CDR1 of SEQ ID NO: 317, aCDR2 of SEQ ID NO: 343 and a CDR3 of SEQ ID NO: 373; (x) an anti-BAFFantibody molecule where the light chain variable domain comprises a CDR1of SEQ ID NO: 256, a CDR2 of SEQ ID NO: 2 and a CDR3 of SEQ ID NO: 299and a heavy chain variable domain comprises a CDR1 of SEQ ID NO: 317, aCDR2 of SEQ ID NO: 344 and a CDR3 of SEQ ID NO: 372; (y) an anti-BAFFantibody molecule where the light chain variable domain comprises a CDR1of SEQ ID NO: 255, a CDR2 of SEQ ID NO: 281 and a CDR3 of SEQ ID NO: 298and a heavy chain variable domain comprises a CDR1 of SEQ ID NO: 318, aCDR2 of SEQ ID NO: 343 and a CDR3 of SEQ ID NO: 374; (z) an anti-BAFFantibody molecule where the light chain variable domain comprises a CDR1of SEQ ID NO: 257, a CDR2 of SEQ ID NO: 282 and a CDR3 of SEQ ID NO: 300and a heavy chain variable domain comprises a CDR1 of SEQ ID NO: 319, aCDR2 of SEQ ID NO: 345 and a CDR3 of SEQ ID NO: 375; (aa) an anti-BAFFantibody molecule where the light chain variable domain comprises a CDR1of SEQ ID NO: 258, a CDR2 of SEQ ID NO: 283 and a CDR3 of SEQ ID NO: 301and a heavy chain variable domain comprises a CDR1 of SEQ ID NO: 320, aCDR2 of SEQ ID NO: 346 and a CDR3 of SEQ ID NO: 376; (bb) an anti-BAFFantibody molecule where the light chain variable domain comprises a CDR1of SEQ ID NO: 259, a CDR2 of SEQ ID NO: 281 and a CDR3 of SEQ ID NO: 298and a heavy chain variable domain comprises a CDR1 of SEQ ID NO: 317, aCDR2 of SEQ ID NO: 347 and a CDR3 of SEQ ID NO: 377; (cc) an anti-BAFFantibody molecule where the light chain variable domain comprises a CDR1of SEQ ID NO: 260, a CDR2 of SEQ ID NO: 284 and a CDR3 of SEQ ID NO: 294and a heavy chain variable domain comprises a CDR1 of SEQ ID NO: 321, aCDR2 of SEQ ID NO: 348 and a CDR3 of SEQ ID NO: 378; (dd) an anti-BAFFantibody molecule where the light chain variable domain comprises a CDR1of SEQ ID NO: 254, a CDR2 of SEQ ID NO: 2 and a CDR3 of SEQ ID NO: 299and a heavy chain variable domain comprises a CDR1 of SEQ ID NO: 322, aCDR2 of SEQ ID NO: 349 and a CDR3 of SEQ ID NO: 372; (ee) an anti-BAFFantibody molecule where the light chain variable domain comprises a CDR1of SEQ ID NO: 261, a CDR2 of SEQ ID NO: 285 and a CDR3 of SEQ ID NO: 294and a heavy chain variable domain comprises a CDR1 of SEQ ID NO: 323, aCDR2 of SEQ ID NO: 350 and a CDR3 of SEQ ID NO: 378; (ff) an anti-BAFFantibody molecule where the light chain variable domain comprises a CDR1of SEQ ID NO: 262, a CDR2 of SEQ ID NO: 286 and a CDR3 of SEQ ID NO: 302and a heavy chain variable domain comprises a CDR1 of SEQ ID NO: 324, aCDR2 of SEQ ID NO: 351 and a CDR3 of SEQ ID NO: 379; (gg) an anti-BAFFantibody molecule where the light chain variable domain comprises a CDR1of SEQ ID NO: 263, a CDR2 of SEQ ID NO: 6 and a CDR3 of SEQ ID NO: 303and a heavy chain variable domain comprises a CDR1 of SEQ ID NO: 317, aCDR2 of SEQ ID NO: 352 and a CDR3 of SEQ ID NO: 380; (hh) an anti-BAFFantibody molecule where the light chain variable domain comprises a CDR1of SEQ ID NO: 264, a CDR2 of SEQ ID NO: 276 and a CDR3 of SEQ ID NO: 304and a heavy chain variable domain comprises a CDR1 of SEQ ID NO: 325, aCDR2 of SEQ ID NO: 353 and a CDR3 of SEQ ID NO: 381; (ii) an anti-BAFFantibody molecule where the light chain variable domain comprises a CDR1of SEQ ID NO: 265, a CDR2 of SEQ ID NO: 287 and a CDR3 of SEQ ID NO: 305and a heavy chain variable domain comprises a CDR1 of SEQ ID NO: 326, aCDR2 of SEQ ID NO: 354 and a CDR3 of SEQ ID NO: 382; (jj) an anti-BAFFantibody molecule where the light chain variable domain comprises a CDR1of SEQ ID NO: 266, a CDR2 of SEQ ID NO: 287 and a CDR3 of SEQ ID NO: 306and a heavy chain variable domain comprises a CDR1 of SEQ ID NO: 326, aCDR2 of SEQ ID NO: 355 and a CDR3 of SEQ ID NO: 383; (kk) an anti-BAFFantibody molecule where the light chain variable domain comprises a CDR1of SEQ ID NO: 267, a CDR2 of SEQ ID NO: 285 and a CDR3 of SEQ ID NO: 294and a heavy chain variable domain comprises a CDR1 of SEQ ID NO: 327, aCDR2 of SEQ ID NO: 356 and a CDR3 of SEQ ID NO: 369; (ll) an anti-BAFFantibody molecule where the light chain variable domain comprises a CDR1of SEQ ID NO: 268, a CDR2 of SEQ ID NO: 276 and a CDR3 of SEQ ID NO: 306and a heavy chain variable domain comprises a CDR1 of SEQ ID NO: 328, aCDR2 of SEQ ID NO: 357 and a CDR3 of SEQ ID NO: 383; (mm) an anti-BAFFantibody molecule where the light chain variable domain comprises a CDR1of SEQ ID NO: 269, a CDR2 of SEQ ID NO: 288 and a CDR3 of SEQ ID NO: 304and a heavy chain variable domain comprises a CDR1 of SEQ ID NO: 329, aCDR2 of SEQ ID NO: 358 and a CDR3 of SEQ ID NO: 384; (nn) an anti-BAFFantibody molecule where the light chain variable domain comprises a CDR1of SEQ ID NO: 270, a CDR2 of SEQ ID NO: 276 and a CDR3 of SEQ ID NO: 306and a heavy chain variable domain comprises a CDR1 of SEQ ID NO: 330, aCDR2 of SEQ ID NO: 359 and a CDR3 of SEQ ID NO: 385; (oo) an anti-BAFFantibody molecule where the light chain variable domain comprises a CDR1of SEQ ID NO: 271, a CDR2 of SEQ ID NO: 289 and a CDR3 of SEQ ID NO: 307and a heavy chain variable domain comprises a CDR1 of SEQ ID NO: 331, aCDR2 of SEQ ID NO: 360 and a CDR3 of SEQ ID NO: 385; (pp) an anti-BAFFantibody molecule where the light chain variable domain comprises a CDR1of SEQ ID NO: 261, a CDR2 of SEQ ID NO: 285 and a CDR3 of SEQ ID NO: 294and a heavy chain variable domain comprises a CDR1 of SEQ ID NO: 332, aCDR2 of SEQ ID NO: 361 and a CDR3 of SEQ ID NO: 386; (qq) an anti-BAFFantibody molecule where the light chain variable domain comprises a CDR1of SEQ ID NO: 272, a CDR2 of SEQ ID NO: 289 and a CDR3 of SEQ ID NO: 307and a heavy chain variable domain comprises a CDR1 of SEQ ID NO: 331, aCDR2 of SEQ ID NO: 362 and a CDR3 of SEQ ID NO: 385; (rr) an anti-BAFFantibody molecule where the light chain variable domain comprises a CDR1of SEQ ID NO: 266, a CDR2 of SEQ ID NO: 287 and a CDR3 of SEQ ID NO: 306and a heavy chain variable domain comprises a CDR1 of SEQ ID NO: 326, aCDR2 of SEQ ID NO: 355 and a CDR3 of SEQ ID NO: 383; (ss) an anti-BAFFantibody molecule where the light chain variable domain comprises a CDR1of SEQ ID NO: 270, a CDR2 of SEQ ID NO: 276 and a CDR3 of SEQ ID NO: 306and a heavy chain variable domain comprises a CDR1 of SEQ ID NO: 330, aCDR2 of SEQ ID NO: 359 and a CDR3 of SEQ ID NO: 285; (tt) an anti-BAFFantibody molecule where the light chain variable domain comprises a CDR1of SEQ ID NO: 270, a CDR2 of SEQ ID NO: 276 and a CDR3 of SEQ ID NO: 306and a heavy chain variable domain comprises a CDR1 of SEQ ID NO: 392, aCDR2 of SEQ ID NO: 363 and a CDR3 of SEQ ID NO: 387; (uu) an anti-BAFFantibody molecule where the light chain variable domain comprises a CDR1of SEQ ID NO: 273, a CDR2 of SEQ ID NO: 276 and a CDR3 of SEQ ID NO: 308and a heavy chain variable domain comprises a CDR1 of SEQ ID NO: 333, aCDR2 of SEQ ID NO: 364 and a CDR3 of SEQ ID NO: 388; (vv) an anti-BAFFantibody molecule where the light chain variable domain comprises a CDR1of SEQ ID NO: 274, a CDR2 of SEQ ID NO: 290 and a CDR3 of SEQ ID NO: 309and a heavy chain variable domain comprises a CDR1 of SEQ ID NO: 334, aCDR2 of SEQ ID NO: 365 and a CDR3 of SEQ ID NO: 389; (ww) an anti-BAFFantibody molecule where the light chain variable domain comprises a CDR1of SEQ ID NO: 275, a CDR2 of SEQ ID NO: 291 and a CDR3 of SEQ ID NO: 310and a heavy chain variable domain comprises a CDR1 of SEQ ID NO: 335, aCDR2 of SEQ ID NO: 366 and a CDR3 of SEQ ID NO: 390; and (xx) ananti-BAFF antibody molecule where the light chain variable domaincomprises a CDR1 of SEQ ID NO: 258, a CDR2 of SEQ ID NO: 292 and a CDR3of SEQ ID NO: 311 and a heavy chain variable domain comprises a CDR1 ofSEQ ID NO: 336, a CDR2 of SEQ ID NO: 367 and a CDR3 of SEQ ID NO: 391.

In another embodiment of the present invention, the anti-BAFF antibodymolecule comprises a light chain variable region of any one of SEQ IDNOS: 82-97, and a heavy chain variable region of any one of SEQ ID NOS:100-115. In a preferred embodiment, the present invention providesmonoclonal antibodies with the combinations of light chain variable andheavy chain variable regions of SEQ ID NO: 82/101, 88/101, 94/112 or93/114. Such variable regions can be combined with human constantregions.

In an additional embodiment of the present invention, the anti-BAFFantibody molecule neutralizes all three forms of human BAFF, the formsof which include membrane bound (mbBAFF), soluble trimeric BAFF, andsoluble 60-mer BAFF. In particular, the anti-BAFF antibody molecules ofthe present invention neutralize human soluble 60-mer BAFF. Furthermore,the anti-BAFF antibody molecules of the present invention neutralizehuman soluble trimeric BAFF. Finally, the anti-BAFF antibody moleculesof the present invention neutralize human membrane-bound BAFF.

In a further embodiment, the present invention relates to an anti-BAFFantibody molecule comprising a humanized light chain variable domaincomprising the CDRs of SEQ ID NO: 76, 16 and 17 and framework regionshaving an amino acid sequence at least 90% identical, at least 93%identical or at least 95% identical to the amino acid sequence of theframework regions of the variable domain light chain amino acid sequenceof SEQ ID NO: 82 and a humanized heavy chain variable domain comprisingthe CDRs of SEQ ID NO: 37, 38 and 39 and framework regions having anamino acid sequence at least 90% identical, at least 93% identical or atleast 95% identical to the amino acid sequence of the framework regionsof the variable domain heavy chain amino acid sequence of SEQ ID NO:101. In one embodiment, the anti-BAFF antibody molecule is a humanizedmonoclonal antibody.

In a further embodiment, the present invention relates to an anti-BAFFantibody molecule comprising a humanized light chain variable domaincomprising the CDRs of SEQ ID NO: 15, 16 and 17 and framework regionshaving an amino acid sequence at least 90% identical, at least 93%identical or at least 95% identical to the amino acid sequence of theframework regions of the variable domain light chain amino acid sequenceof SEQ ID NO: 88 and a humanized heavy chain variable domain comprisingthe CDRs of SEQ ID NO: 37, 38 and 39 and framework regions having anamino acid sequence at least 90% identical, at least 93% identical or atleast 95% identical to the amino acid sequence of the framework regionsof the variable domain heavy chain amino acid sequence of SEQ ID NO:101. In one embodiment, the anti-BAFF antibody molecule is a humanizedmonoclonal antibody.

In a further embodiment, the present invention relates to an anti-BAFFantibody molecule comprising a humanized light chain variable domaincomprising the CDRs of SEQ ID NO:5, 8 and 9 and framework regions havingan amino acid sequence at least 90% identical, at least 93% identical orat least 95% identical to the amino acid sequence of the frameworkregions of the variable domain light chain amino acid sequence of SEQ IDNO: 94 and a humanized heavy chain variable domain comprising the CDRsof SEQ ID NO: 81, 29 and 30 and framework regions having an amino acidsequence at least 90% identical, at least 93% identical or at least 95%identical to the amino acid sequence of the framework regions of thevariable domain heavy chain amino acid sequence of SEQ ID NO: 112. Inone embodiment, the anti-BAFF antibody molecule is a humanizedmonoclonal antibody.

In a further embodiment, the present invention relates to an anti-BAFFantibody molecule comprising a humanized light chain variable domaincomprising the CDRs of SEQ ID NO: 5, 8 and 9 and framework regionshaving an amino acid sequence at least 90% identical, at least 93%identical or at least 95% identical to the amino acid sequence of theframework regions of the variable domain light chain amino acid sequenceof SEQ ID NO: 93 and a humanized heavy chain variable domain comprisingthe CDRs of SEQ ID NO: 81, 29 and 30 and framework regions having anamino acid sequence at least 90% identical, at least 93% identical or atleast 95% identical to the amino acid sequence of the framework regionsof the variable domain heavy chain amino acid sequence of SEQ ID NO:114. In one embodiment, the anti-BAFF antibody molecule is a humanizedmonoclonal antibody.

In specific embodiments, it is contemplated that chimeric antibodieswith switched CDR regions (i.e., for example switching one or two CDRsof one of the mouse antibodies or humanized antibody derived therefromwith the analogous CDR from another mouse antibody or humanized antibodyderived therefrom) between these exemplary immunoglobulins may yielduseful antibodies.

In certain embodiments, the humanized anti-BAFF antibody is an antibodyfragment. Various antibody fragments have been generally discussed aboveand there are techniques that have been developed for the production ofantibody fragments. Fragments can be derived via proteolytic digestionof intact antibodies (see, e.g., Morimoto et al., 1992, Journal ofBiochemical and Biophysical Methods 24:107-117; and Brennan et al.,1985, Science 229:81). Alternatively, the fragments can be produceddirectly in recombinant host cells. For example, Fab′-SH fragments canbe directly recovered from E. coli and chemically coupled to formF(ab′)₂ fragments (see, e.g., Carter et al., 1992, Bio/Technology10:163-167). By another approach, F(ab′)₂ fragments can be isolateddirectly from recombinant host cell culture. Other techniques for theproduction of antibody fragments will be apparent to the skilledpractitioner. Accordingly, in one aspect, the present invention providesantibody fragments comprising the CDRs described herein, in particularone of the combinations of L-CDR1, L-CDR2, L-CDR3, H-CDR1, H-CDR2 andH-CDR3 described herein. In a further aspect, the present inventionprovides antibody fragments comprising the variable regions describedherein, for example one of the combinations of light chain variableregions and heavy chain variable regions described herein.

In some embodiments, the antibody or antibody fragment includes aconstant region that mediates effector function. The constant region canprovide antibody-dependent cellular cytotoxicity (ADCC),antibody-dependent cellular phagocytosis (ADCP) and/orcomplement-dependent cytotoxicity (CDC) responses against a BAFFexpressing target cell. The effector domain(s) can be, for example, anFc region of an Ig molecule.

The effector domain of an antibody can be from any suitable vertebrateanimal species and isotypes. The isotypes from different animal speciesdiffer in the abilities to mediate effector functions. For example, theability of human immunoglobulin to mediate CDC and ADCC/ADCP isgenerally in the order of IgM≈IgG₁≈IgG₃>IgG₂>IgG₄ andIgG₁≈IgG₃>IgG₂/IgM/IgG₄, respectively. Murine immunoglobulins mediateCDC and ADCC/ADCP generally in the order of murineIgM≈IgG₃>>IgG_(2b)>IgG_(2a)>>IgG₁ and IgG_(2b)>IgG_(2a)>IgG₁>>IgG₃,respectively. In another example, murine IgG_(2a) mediates ADCC whileboth murine IgG_(2a) and IgM mediate CDC.

Antibody Modifications

The humanized anti-BAFF antibodies and agents can include modificationsof the humanized anti-BAFF antibody or antigen-binding fragment thereof.For example, it may be desirable to modify the antibody with respect toeffector function, so as to enhance the effectiveness of the antibody intreating cancer. One such modification is the introduction of cysteineresidue(s) into the Fc region, thereby allowing interchain disulfidebond formation in this region. The homodimeric antibody thus generatedcan have improved internalization capability and/or increasedcomplement-mediated cell killing and/or antibody-dependent cellularcytotoxicity (ADCC). See, for example, Caron et al., 1992, J. Exp Med.176:1191-1195; and Shopes, 1992, J. Immunol. 148:2918-2922. Homodimericantibodies having enhanced anti-tumor activity can also be preparedusing heterobifunctional cross-linkers as described in Wolff et al.,1993, Cancer Research 53: 2560-2565. Alternatively, an antibody can beengineered to contain dual Fc regions, enhancing complement lysis andADCC capabilities of the antibody. See Stevenson et al., 1989,Anti-Cancer Drug Design 3: 219-230.

Antibodies with improved ability to support ADCC have been generated bymodifying the glycosylation pattern of their Fc region. This is possiblesince antibody glycosylation at the asparagine residue, N297, in theC_(H2) domain is involved in the interaction between IgG and Fcγreceptors prerequisite to ADCC. Host cell lines have been engineered toexpress antibodies with altered glycosylation, such as increasedbisecting N-acetylglucosamine or reduced fucose. Fucose reductionprovides greater enhancement to ADCC activity than does increasing thepresence of bisecting N-acetylglucosamine. Moreover, enhancement of ADCCby low fucose antibodies is independent of the FcγRIIIa V/Fpolymorphism.

Modifying the amino acid sequence of the Fc region of antibodies is analternative to glycosylation engineering to enhance ADCC. The bindingsite on human IgG₁ for Fcγ receptors has been determined by extensivemutational analysis. This led to the generation of humanized IgG₁antibodies with Fc mutations that increase the binding affinity forFcγRIIIa and enhance ADCC in vitro. Additionally, Fc variants have beenobtained with many different permutations of binding properties, e.g.,improved binding to specific FcγR receptors with unchanged or diminishedbinding to other FcγR receptors.

Another aspect includes immunoconjugates comprising the humanizedantibody or fragments thereof conjugated to a cytotoxic agent such as achemotherapeutic agent, a toxin (e.g., an enzymatically active toxin ofbacterial, fungal, plant, or animal origin, or fragments thereof), or aradioactive isotope (i.e., a radioconjugate).

Chemotherapeutic agents useful in the generation of suchimmunoconjugates have been described above. Enzymatically active toxinsand fragments thereof that can be used to form useful immunoconjugatesinclude diphtheria A chain, nonbinding active fragments of diphtheriatoxin, exotoxin A chain (from Pseudomonas aeruginosa), ricin A chain,abrin A chain, modeccin A chain, alpha-sarcin, Aleurites fordiiproteins, dianthin proteins, Phytolaca americana proteins (PAPI, PAPII,and PAP-S), Momordica charantia inhibitor, curcin, crotin, Sapaonariaofficinalis inhibitor, gelonin, mitogellin, restrictocin, phenomycin,enomycin, the tricothecenes, and the like. A variety of radionuclidesare available for the production of radioconjugated humanized anti-BAFFantibodies. Examples include ²¹²Bi, ¹³¹I, ¹³¹In, ⁹⁰Y and ¹⁸⁶Re.

Conjugates of the humanized anti-BAFF antibody and cytotoxic orchemotherapeutic agent can be made by known methods, using a variety ofbifunctional protein coupling agents such asN-succinimidyl-3-(2-pyridyldithiol) propionate (SPDP), iminothiolane(IT), bifunctional derivatives of imidoesters (such as dimethyladipimidate HCL), active esters (such as disuccinimidyl suberate),aldehydes (such as glutareldehyde), bis-azido compounds (such asbis(p-azidobenzoyl) hexanediamine), bis-diazonium derivatives (such asbis-(p-diazoniumbenzoyl)-ethylenediamine), diisocyanates (such astoluene 2,6-diisocyanate), and bis-active fluorine compounds (such as1,5-difluoro-2,4-dinitrobenzene). For example, a ricin immunotoxin canbe prepared as described in Vitetta et al., 1987, Science 238:1098.Carbon-14-labeled 1-isothiocyanatobenzyl-3-methyldiethylenetriaminepentaacetic acid (MX-DTPA) is an exemplary chelating agent forconjugation of radionucleotide to the antibody. Conjugates also can beformed with a cleavable linker.

The humanized anti-BAFF antibodies disclosed herein can also beformulated as immunoliposomes. Liposomes containing the antibody areprepared by methods known in the art, such as described in Epstein etal., 1985, Proc. Natl. Acad. Sci. USA 82:3688; Hwang et al., 1980, Proc.Natl. Acad. Sci. USA 77:4030; and U.S. Pat. Nos. 4,485,045 and4,544,545. Liposomes having enhanced circulation time are disclosed, forexample, in U.S. Pat. No. 5,013,556.

Particularly useful liposomes can be generated by the reverse phaseevaporation method with a lipid composition comprisingphosphatidylcholine, cholesterol and PEG-derivatizedphosphatidylethanolamine (PEG-PE). Liposomes are extruded throughfilters of defined pore size to yield liposomes with the desireddiameter. Fab′ fragments of an antibody disclosed herein can beconjugated to the liposomes as described in Martin et al., 1982, J.Biol. Chem. 257:286-288 via a disulfide interchange reaction. Achemotherapeutic agent (such as doxorubicin) is optionally containedwithin the liposome. See, e.g., Gabizon et al., 1989, J. National CancerInst. 81(19):1484.

The antibodies described and disclosed herein can also be used in ADEPT(Antibody-Directed Enzyme Prodrug Therapy) procedures by conjugating theantibody to a prodrug-activating enzyme that converts a prodrug (e.g., apeptidyl chemotherapeutic agent), to an active anti-cancer drug. See,for example, WO 81/01145, WO 88/07378, and U.S. Pat. No. 4,975,278. Theenzyme component of the immunoconjugate useful for ADEPT is an enzymecapable of acting on a prodrug in such a way so as to convert it intoits more active, cytotoxic form. Specific enzymes that are useful inADEPT include, but are not limited to, alkaline phosphatase forconverting phosphate-containing prodrugs into free drugs; arylsulfatasefor converting sulfate-containing prodrugs into free drugs; cytosinedeaminase for converting non-toxic 5-fluorocytosine into the anti-cancerdrug, 5-fluorouracil; proteases, such as serratia protease, thermolysin,subtilisin, carboxypeptidases, and cathepsins (such as cathepsins B andL), for converting peptide-containing prodrugs into free drugs;D-alanylcarboxypeptidases, for converting prodrugs containing D-aminoacid substituents; carbohydrate-cleaving enzymes such as β-galactosidaseand neuraminidase for converting glycosylated prodrugs into free drugs;β-lactamase for converting drugs derivatized with β-lactams into freedrugs; and penicillin amidases, such as penicillin V amidase orpenicillin G amidase, for converting drugs derivatized at their aminenitrogens with phenoxyacetyl or phenylacetyl groups, respectively, intofree drugs. Alternatively, antibodies having enzymatic activity(“abzymes”) can be used to convert the prodrugs into free active drugs(see, for example, Massey, 1987, Nature 328: 457-458). Antibody-abzymeconjugates can be prepared by known methods for delivery of the abzymeto a tumor cell population, for example, by covalently binding theenzyme to the humanized anti-BAFF antibody/heterobifunctionalcrosslinking reagents discussed above. Alternatively, fusion proteinscomprising at least the antigen binding region of an antibody disclosedherein linked to at least a functionally active portion of an enzyme asdescribed above can be constructed using recombinant DNA techniques(see, e.g., Neuberger et al., 1984, Nature 312:604-608).

In certain embodiments, it may be desirable to use a humanized anti-BAFFantibody fragment, rather than an intact antibody, to increase tissuepenetration, for example. It may be desirable to modify the antibodyfragment in order to increase its serum half-life. This can be achieved,for example, by incorporation of a salvage receptor binding epitope intothe antibody fragment. In one method, the appropriate region of theantibody fragment can be altered (e.g., mutated), or the epitope can beincorporated into a peptide tag that is then fused to the antibodyfragment at either end or in the middle, for example, by DNA or peptidesynthesis. See, e.g., WO 96/32478.

In other embodiments, covalent modifications of the humanized anti-BAFFantibody are also included. Covalent modifications include modificationof cysteinyl residues, histidyl residues, lysinyl and amino-terminalresidues, arginyl residues, tyrosyl residues, carboxyl side groups(aspartyl or glutamyl), glutaminyl and asparaginyl residues, or seryl,or threonyl residues. Another type of covalent modification involveschemically or enzymatically coupling glycosides to the antibody. Suchmodifications may be made by chemical synthesis or by enzymatic orchemical cleavage of the antibody, if applicable. Other types ofcovalent modifications of the antibody can be introduced into themolecule by reacting targeted amino acid residues of the antibody withan organic derivatizing agent that is capable of reacting with selectedside chains or the amino- or carboxy-terminal residues.

Removal of any carbohydrate moieties present on the antibody can beaccomplished chemically or enzymatically. Chemical deglycosylation isdescribed by Hakimuddin et al., 1987, Arch. Biochem. Biophys. 259:52 andby Edge et al., 1981, Anal. Biochem., 118:131. Enzymatic cleavage ofcarbohydrate moieties on antibodies can be achieved by the use of avariety of endo- and exo-glycosidases as described by Thotakura et al.,1987, Meth. Enzymol 138:350.

Another type of useful covalent modification comprises linking theantibody to one of a variety of nonproteinaceous polymers, e.g.,polyethylene glycol, polypropylene glycol, or polyoxyalkylenes, in themanner set forth in one or more of U.S. Pat. No. 4,640,835, U.S. Pat.No. 4,496,689, U.S. Pat. No. 4,301,144, U.S. Pat. No. 4,670,417, U.S.Pat. No. 4,791,192 and U.S. Pat. No. 4,179,337.

Humanization and Amino Acid Sequence Variants

Amino acid sequence variants of the anti-BAFF antibody can be preparedby introducing appropriate nucleotide changes into the anti-BAFFantibody DNA, or by peptide synthesis. Such variants include, forexample, deletions from, and/or insertions into and/or substitutions of,residues within the amino acid sequences of the anti-BAFF antibodies ofthe examples herein. Any combination of deletions, insertions, andsubstitutions is made to arrive at the final construct, provided thatthe final construct possesses the desired characteristics. The aminoacid changes also may alter post-translational processes of thehumanized or variant anti-BAFF antibody, such as changing the number orposition of glycosylation sites.

A useful method for identification of certain residues or regions of theanti-BAFF antibody that are preferred locations for mutagenesis iscalled “alanine scanning mutagenesis,” as described by Cunningham andWells (Science, 244:1081-1085 (1989)). Here, a residue or group oftarget residues are identified (e.g., charged residues such as arg, asp,his, lys, and glu) and replaced by a neutral or negatively charged aminoacid (typically alanine) to affect the interaction of the amino acidswith BAFF antigen. Those amino acid locations demonstrating functionalsensitivity to the substitutions then are refined by introducing furtheror other variants at, or for, the sites of substitution. Thus, while thesite for introducing an amino acid sequence variation is predetermined,the nature of the mutation per se need not be predetermined. Forexample, to analyze the performance of a mutation at a given site,alanine scanning or random mutagenesis is conducted at the target codonor region and the expressed anti-BAFF antibody variants are screened forthe desired activity.

Amino acid sequence insertions include amino- and/or carboxyl-terminalfusions ranging in length from one residue to polypeptides containing ahundred or more residues, as well as intrasequence insertions of singleor multiple amino acid residues. Examples of terminal insertions includean anti-BAFF antibody fused to an epitope tag. Other insertionalvariants of the anti-BAFF antibody molecule include a fusion to the N-or C-terminus of the anti-BAFF antibody of an enzyme or a polypeptidewhich increases the serum half-life of the antibody.

Another type of variant is an amino acid substitution variant. Thesevariants have at least one amino acid residue in the anti-BAFF antibodymolecule removed and a different residue inserted in its place. Thesites of greatest interest for substitutional mutagenesis include thehypervariable regions, but FR alterations are also contemplated.Conservative substitutions are shown below under the heading of“preferred substitutions”. If such substitutions result in a change inbiological activity, then more substantial changes, denominated“exemplary substitutions”, or as further described below in reference toamino acid classes, may be introduced and the products screened.

Original Residue Exemplary Substitutions Preferred Substitutions Ala (A)val; leu; ile val Arg (R) lys; gln; asn lys Asn (N) gln; his; asp, lys;arg gln Asp (D) glu; asn glu Cys (C) ser; ala ser Gln (Q) asn; glu asnGlu (E) asp; gln asp Gly (G) ala ala His (H) arg; asn; gln; lys; arg Ile(I) leu; val; met; ala; phe; norleucine leu Leu (L) ile; norleucine;val; met; ala; phe ile Lys (K) arg; gln; asn arg Met (M) leu; phe; ileleu Phe (F) tyr; leu; val; ile; ala; tyr Pro (P) ala ala Ser (S) thr thrThr (T) ser ser Trp (W) tyr; phe tyr Tyr (Y) phe; trp; thr; ser phe Val(V) leu; ile; met; phe ala; norleucine; leu

In protein chemistry, it is generally accepted that the biologicalproperties of the antibody can be accomplished by selectingsubstitutions that differ significantly in their effect on maintaining(a) the structure of the polypeptide backbone in the area of thesubstitution, for example, as a sheet or helical conformation, (b) thecharge or hydrophobicity of the molecule at the target site, or (c) thebulk of the side chain. Naturally occurring residues are divided intogroups based on common side-chain properties:

(1) hydrophobic: norleucine, met, ala, val, leu, ile;(2) neutral hydrophilic: cys, ser, thr;(3) acidic: asp, glu;(4) basic: asn, gin, his, lys, arg;(5) residues that influence chain orientation: gly, pro; and(6) aromatic: trp, tyr, phe.

Non-conservative substitutions will entail exchanging a member of one ofthese classes for another class.

Any cysteine residue not involved in maintaining the proper conformationof the humanized or variant anti-BAFF antibody also may be substituted,generally with serine, to improve the oxidative stability of themolecule, prevent aberrant crosslinking, or provide for establishedpoints of conjugation to a cytotoxic or cytostatic compound. Conversely,cysteine bond(s) may be added to the antibody to improve its stability(particularly where the antibody is an antibody fragment such as an Fvfragment).

A type of substitutional variant involves substituting one or morehypervariable region residues of a parent antibody (e.g., a humanized orhuman antibody). Generally, the resulting variant(s) selected forfurther development will have improved biological properties relative tothe parent antibody from which they are generated. A convenient way forgenerating such substitutional variants is affinity maturation usingphage display. Briefly, several hypervariable region sites (e.g., 6-7sites) are mutated to generate all possible amino substitutions at eachsite. The antibody variants thus generated are displayed in a monovalentfashion from filamentous phage particles as fusions to the gene IIIproduct of M13 packaged within each particle. The phage-displayedvariants are then screened for their biological activity (e.g., bindingaffinity). In order to identify candidate hypervariable region sites formodification, alanine scanning mutagenesis can be performed to identifyhypervariable region residues contributing significantly to antigenbinding. Alternatively, or in addition, it may be beneficial to analyzea crystal structure of the antigen-antibody complex to identify contactpoints between the antibody and human BAFF. Such contact residues andneighboring residues are candidates for substitution according to thetechniques elaborated herein. Once such variants are generated, thepanel of variants is subjected to screening as described herein andantibodies with superior properties in one or more relevant assays maybe selected for further development.

Another type of amino acid variant of the antibody alters the originalglycosylation pattern of the antibody. By “altering” is meant deletingone or more carbohydrate moieties found in the antibody, and/or addingone or more glycosylation sites that are not present in the antibody. Insome embodiments, it may be desirable to modify the antibodies of theinvention to add glycosylations sites. Glycosylation of antibodies istypically either N-linked or O-linked. N-linked refers to the attachmentof the carbohydrate moiety to the side chain of an asparagine residue.The tripeptide sequences asparagine-X-serine and asparagine-X-threonine,where X is any amino acid except proline, are the recognition sequencesfor enzymatic attachment of the carbohydrate moiety to the asparagineside chain. Thus, the presence of either of these tripeptide sequencesin a polypeptide creates a potential glycosylation site. O-linkedglycosylation refers to the attachment of one of the sugarsN-aceylgalactosamine, galactose, or xylose to a hydroxyamino acid, mostcommonly serine or threonine, although 5-hydroxyproline or5-hydroxylysine may also be used. Thus, in order to glycosylate a givenprotein, e.g., an antibody, the amino acid sequence of the protein isengineered to contain one or more of the above-described tripeptidesequences (for N-linked glycosylation sites). The alteration may also bemade by the addition of, or substitution by, one or more serine orthreonine residues to the sequence of the original antibody (forO-linked glycosylation sites).

Nucleic acid molecules encoding amino acid sequence variants of theanti-BAFF antibody are prepared by a variety of methods known in theart. These methods include, but are not limited to, isolation from anatural source (in the case of naturally occurring amino acid sequencevariants) or preparation by oligonucleotide-mediated (or site-directed)mutagenesis, PCR mutagenesis, and cassette mutagenesis of an earlierprepared variant or a non-variant version of the anti-BAFF antibody.

Polynucleotides, Vectors, Host Cells, and Recombinant Methods

Other embodiments encompass isolated polynucleotides that comprise asequence encoding a humanized anti-BAFF antibody, vectors, and hostcells comprising the polynucleotides, and recombinant techniques forproduction of the humanized antibody. The isolated polynucleotides canencode any desired form of the anti-BAFF antibody including, forexample, full length monoclonal antibodies, Fab, Fab′, F(ab′)₂, and Fvfragments, diabodies, linear antibodies, single-chain antibodymolecules, and multispecific antibodies formed from antibody fragments.

In one embodiment, the present invention provides isolatedpolynucleotides comprising combinations of light chain variable andheavy chain variable regions of SEQ ID NO: 40/58, 42/60, 44/62, 46/64,48/66, 50/68, 52/70, 54/72, 56/74, 118/182, 120/184, 122/186, 124/188,126/190, 128/192, 130/194, 132/196, 134/198, 136/200, 138/202, 140/204,142/206, 144/208, 146/210, 148/212, 150/214, 152/216, 154/218, 156/220,158/222, 160/224, 162/226, 164/228, 166/230, 168/232, 170/236, 172/237,172/239, 174/241, 176/243, 178/245 or 180/247.

Some embodiments include isolated polynucleotides comprising sequencesthat encode the light chain variable region of an antibody or antibodyfragment having the amino acid sequence of any of SEQ ID NOs: 82, 83,84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, or 97. Exemplarypolynucleotide sequences encoding such amino acid sequences are SEQ IDNOs: 234, 392, 393 and 394. Other embodiments include isolatedpolynucleotides comprising sequences that encode the heavy chainvariable region of an antibody or antibody fragment having the aminoacid sequence of any of the SEQ ID NOs: 100, 101, 102, 103, 104, 105,106, 107, 108, 109, 110, 111, 112, 113, 114 or 115. Exemplarypolynucleotide sequences encoding such amino acid sequences are SEQ IDNOs: 395, 396 and 397.

In one embodiment, the isolated polynucleotide comprises a light chainvariable region is SEQ ID NO: 234 and the heavy chain variable region isSEQ ID NO: 396, the light chain variable region is SEQ ID NO: 393 andthe heavy chain variable region is SEQ ID NO: 396, the light chainvariable region is SEQ ID NO: 395 and the heavy chain variable region isSEQ ID NO: 397 or the light chain variable region is SEQ ID NO: 394 andthe heavy chain variable region is SEQ ID NO: 398.

The polynucleotide(s) that comprise a sequence encoding a humanizedanti-BAFF antibody or a fragment or chain thereof can be fused to one ormore regulatory or control sequence, as known in the art, and can becontained in suitable expression vectors or host cell as known in theart. Each of the polynucleotide molecules encoding the heavy or lightchain variable domains can be independently fused to a polynucleotidesequence encoding a constant domain, such as a human constant domain,enabling the production of intact antibodies. Alternatively,polynucleotides, or portions thereof, can be fused together, providing atemplate for production of a single chain antibody.

For recombinant production, a polynucleotide encoding the antibody isinserted into a replicable vector for cloning (amplification of the DNA)or for expression. Many suitable vectors for expressing the recombinantantibody are available. The vector components generally include, but arenot limited to, one or more of the following: a signal sequence, anorigin of replication, one or more marker genes, an enhancer element, apromoter, and a transcription termination sequence.

The humanized anti-BAFF antibodies can also be produced as fusionpolypeptides, in which the antibody is fused with a heterologouspolypeptide, such as a signal sequence or other polypeptide having aspecific cleavage site at the amino terminus of the mature protein orpolypeptide. The heterologous signal sequence selected is typically onethat is recognized and processed (i.e., cleaved by a signal peptidase)by the host cell. For prokaryotic host cells that do not recognize andprocess the humanized anti-BAFF antibody signal sequence, the signalsequence can be substituted by a prokaryotic signal sequence. The signalsequence can be, for example, alkaline phosphatase, penicillinase,lipoprotein, heat-stable enterotoxin II leaders, and the like. For yeastsecretion, the native signal sequence can be substituted, for example,with a leader sequence obtained from yeast invertase alpha-factor(including Saccharomyces and Kluyveromyces α-factor leaders), acidphosphatase, C. albicans glucoamylase, or the signal described inWO90/13646. In mammalian cells, mammalian signal sequences as well asviral secretory leaders, for example, the herpes simplex gD signal, canbe used. The DNA for such precursor region is ligated in reading frameto DNA encoding the humanized anti-BAFF antibody.

Expression and cloning vectors contain a nucleic acid sequence thatenables the vector to replicate in one or more selected host cells.Generally, in cloning vectors this sequence is one that enables thevector to replicate independently of the host chromosomal DNA, andincludes origins of replication or autonomously replicating sequences.Such sequences are well known for a variety of bacteria, yeast, andviruses. The origin of replication from the plasmid pBR322 is suitablefor most Gram-negative bacteria, the 2-ν. plasmid origin is suitable foryeast, and various viral origins (SV40, polyoma, adenovirus, VSV, andBPV) are useful for cloning vectors in mammalian cells. Generally, theorigin of replication component is not needed for mammalian expressionvectors (the SV40 origin may typically be used only because it containsthe early promoter).

Expression and cloning vectors may contain a gene that encodes aselectable marker to facilitate identification of expression. Typicalselectable marker genes encode proteins that confer resistance toantibiotics or other toxins, e.g., ampicillin, neomycin, methotrexate,or tetracycline, or alternatively, are complement auxotrophicdeficiencies, or in other alternatives supply specific nutrients thatare not present in complex media, e.g., the gene encoding D-alanineracemase for Bacilli.

One example of a selection scheme utilizes a drug to arrest growth of ahost cell. Those cells that are successfully transformed with aheterologous gene produce a protein conferring drug resistance and thussurvive the selection regimen. Examples of such dominant selection usethe drugs neomycin, mycophenolic acid, and hygromycin. Common selectablemarkers for mammalian cells are those that enable the identification ofcells competent to take up a nucleic acid encoding a humanized anti-BAFFantibody, such as DHFR (dihydrofolate reductase), thymidine kinase,metallothionein-I and -II (such as primate metallothionein genes),adenosine deaminase, ornithine decarboxylase, and the like. Cellstransformed with the DHFR selection gene are first identified byculturing all of the transformants in a culture medium that containsmethotrexate (Mtx), a competitive antagonist of DHFR. An appropriatehost cell when wild-type DHFR is employed is the Chinese hamster ovary(CHO) cell line deficient in DHFR activity (e.g., DG44).

Alternatively, host cells (particularly wild-type hosts that containendogenous DHFR) transformed or co-transformed with DNA sequencesencoding anti-BAFF antibody, wild-type DHFR protein, and anotherselectable marker such as aminoglycoside 3′-phosphotransferase (APH),can be selected by cell growth in medium containing a selection agentfor the selectable marker such as an aminoglycosidic antibiotic, e.g.,kanamycin, neomycin, or G418. See, e.g., U.S. Pat. No. 4,965,199.

Where the recombinant production is performed in a yeast cell as a hostcell, the TRP1 gene present in the yeast plasmid YRp7 (Stinchcomb etal., 1979, Nature 282: 39) can be used as a selectable marker. The TRP1gene provides a selection marker for a mutant strain of yeast lackingthe ability to grow in tryptophan, for example, ATCC No. 44076 or PEP4-1(Jones, 1977, Genetics 85:12). The presence of the trp1 lesion in theyeast host cell genome then provides an effective environment fordetecting transformation by growth in the absence of tryptophan.Similarly, Leu2p-deficient yeast strains such as ATCC 20,622 and 38,626are complemented by known plasmids bearing the LEU2 gene.

In addition, vectors derived from the 1.6 μm circular plasmid pKD1 canbe used for transformation of Kluyveromyces yeasts. Alternatively, anexpression system for large-scale production of recombinant calfchymosin was reported for K. lactis (Van den Berg, 1990, Bio/Technology8:135). Stable multi-copy expression vectors for secretion of maturerecombinant human serum albumin by industrial strains of Kluyveromyceshave also been disclosed (Fleer et al., 1991, Bio/Technology 9:968-975).

Expression and cloning vectors usually contain a promoter that isrecognized by the host organism and is operably linked to the nucleicacid molecule encoding an anti-BAFF antibody or polypeptide chainthereof. Promoters suitable for use with prokaryotic hosts include phoApromoter, β-lactamase and lactose promoter systems, alkalinephosphatase, tryptophan (trp) promoter system, and hybrid promoters suchas the tac promoter. Other known bacterial promoters are also suitable.Promoters for use in bacterial systems also will contain aShine-Dalgarno (S.D.) sequence operably linked to the DNA encoding thehumanized anti-BAFF antibody.

Many eukaryotic promoter sequences are known. Virtually all eukaryoticgenes have an AT-rich region located approximately 25 to 30 basesupstream from the site where transcription is initiated. Anothersequence found 70 to 80 bases upstream from the start of transcriptionof many genes is a CNCAAT region where N may be any nucleotide. At the3′ end of most eukaryotic genes is an AATAAA sequence that may be thesignal for addition of the poly A tail to the 3′ end of the codingsequence. All of these sequences are suitably inserted into eukaryoticexpression vectors.

Examples of suitable promoting sequences for use with yeast hostsinclude the promoters for 3-phosphoglycerate kinase or other glycolyticenzymes, such as enolase, glyceraldehyde-3-phosphate dehydrogenase,hexokinase, pyruvate decarboxylase, phosphofructokinase,glucose-6-phosphate isomerase, 3-phosphoglycerate mutase, pyruvatekinase, triosephosphate isomerase, phosphoglucose isomerase, andglucokinase.

Inducible promoters have the additional advantage of transcriptioncontrolled by growth conditions. These include yeast promoter regionsfor alcohol dehydrogenase 2, isocytochrome C, acid phosphatase,derivative enzymes associated with nitrogen metabolism, metallothionein,glyceraldehyde-3-phosphate dehydrogenase, and enzymes responsible formaltose and galactose utilization. Suitable vectors and promoters foruse in yeast expression are further described in EP 73,657. Yeastenhancers also are advantageously used with yeast promoters.

Humanized anti-BAFF antibody transcription from vectors in mammalianhost cells is controlled, for example, by promoters obtained from thegenomes of viruses such as polyoma virus, fowlpox virus, adenovirus(such as Adenovirus 2), bovine papilloma virus, avian sarcoma virus,cytomegalovirus, a retrovirus, hepatitis-B virus and Simian Virus 40(SV40), from heterologous mammalian promoters, e.g., the actin promoteror an immunoglobulin promoter, or from heat-shock promoters, providedsuch promoters are compatible with the host cell systems.

The early and late promoters of the SV40 virus are conveniently obtainedas an SV40 restriction fragment that also contains the SV40 viral originof replication. The immediate early promoter of the humancytomegalovirus is conveniently obtained as a HindIII E restrictionfragment. A system for expressing DNA in mammalian hosts using thebovine papilloma virus as a vector is disclosed in U.S. Pat. No.4,419,446. A modification of this system is described in U.S. Pat. No.4,601,978. See also Reyes et al., 1982, Nature 297:598-601, disclosingexpression of human p-interferon cDNA in mouse cells under the controlof a thymidine kinase promoter from herpes simplex virus. Alternatively,the Rous sarcoma virus long terminal repeat can be used as the promoter.

Another useful element that can be used in a recombinant expressionvector is an enhancer sequence, which is used to increase thetranscription of a DNA encoding a humanized anti-BAFF antibody by highereukaryotes. Many enhancer sequences are now known from mammalian genes(e.g., globin, elastase, albumin, α-fetoprotein, and insulin).Typically, however, an enhancer from a eukaryotic cell virus is used.Examples include the SV40 enhancer on the late side of the replicationorigin (bp 100-270), the cytomegalovirus early promoter enhancer, thepolyoma enhancer on the late side of the replication origin, andadenovirus enhancers. See also Yaniv, 1982, Nature 297:17-18 for adescription of enhancing elements for activation of eukaryoticpromoters. The enhancer may be spliced into the vector at a position 5′or 3′ to the humanized anti-BAFF antibody-encoding sequence, but ispreferably located at a site 5′ from the promoter.

Expression vectors used in eukaryotic host cells (yeast, fungi, insect,plant, animal, human, or nucleated cells from other multicellularorganisms) can also contain sequences necessary for the termination oftranscription and for stabilizing the mRNA. Such sequences are commonlyavailable from the 5′ and, occasionally 3′, untranslated regions ofeukaryotic or viral DNAs or cDNAs. These regions contain nucleotidesegments transcribed as polyadenylated fragments in the untranslatedportion of the mRNA encoding anti-BAFF antibody. One usefultranscription termination component is the bovine growth hormonepolyadenylation region. See WO94/11026 and the expression vectordisclosed therein. In some embodiments, humanized anti-BAFF antibodiescan be expressed using the CHEF system. (See, e.g., U.S. Pat. No.5,888,809; the disclosure of which is incorporated by reference herein.)

Suitable host cells for cloning or expressing the DNA in the vectorsherein are the prokaryote, yeast, or higher eukaryote cells describedabove. Suitable prokaryotes for this purpose include eubacteria, such asGram-negative or Gram-positive organisms, for example,Enterobacteriaceae such as Escherichia, e.g., E. coli, Enterobacter,Erwinia, Klebsiella, Proteus, Salmonella, e.g., Salmonella typhimurium,Serratia, e.g., Serratia marcescans, and Shigella, as well as Bacillisuch as B. subtilis and B. licheniformis (e.g., B. licheniformis 41 Pdisclosed in DD 266,710 published Apr. 12, 1989), Pseudomonas such as P.aeruginosa, and Streptomyces. One preferred E. coli cloning host is E.coli 294 (ATCC 31,446), although other strains such as E. coli B, E.coli X1776 (ATCC 31,537), and E. coli W3110 (ATCC 27,325) are suitable.These examples are illustrative rather than limiting.

In addition to prokaryotes, eukaryotic microbes such as filamentousfungi or yeast are suitable cloning or expression hosts for humanizedanti-BAFFantibody-encoding vectors. Saccharomyces cerevisiae, or commonbaker's yeast, is the most commonly used among lower eukaryotic hostmicroorganisms. However, a number of other genera, species, and strainsare commonly available and useful herein, such as Schizosaccharomycespombe; Kluyveromyces hosts such as, e.g., K. lactis, K. fragilis (ATCC12,424), K. bulgaricus (ATCC 16,045), K. wickeramii (ATCC 24,178), K.waltii (ATCC 56,500), K. drosophilarum (ATCC 36,906), K. thermotolerans,and K. marxianus; yarrowia (EP 402,226); Pichia pastors (EP 183,070);Candida; Trichoderma reesia (EP 244,234); Neurospora crassa;Schwanniomyces such as Schwanniomyces occidentalis; and filamentousfungi such as, e.g., Neurospora, Penicillium, Tolypocladium, andAspergillus hosts such as A. nidulans and A. niger.

Suitable host cells for the expression of glycosylated humanizedanti-BAFF antibody are derived from multicellular organisms. Examples ofinvertebrate cells include plant and insect cells, including, e.g.,numerous baculoviral strains and variants and corresponding permissiveinsect host cells from hosts such as Spodoptera frugiperda(caterpillar), Aedes aegypti (mosquito), Aedes albopictus (mosquito),Drosophila melanogaster (fruitfly), and Bombyx mori (silk worm). Avariety of viral strains for transfection are publicly available, e.g.,the L-1 variant of Autographa californica NPV and the Bm-5 strain ofBombyx mori NPV, and such viruses may be used, particularly fortransfection of Spodoptera frugiperda cells.

Plant cell cultures of cotton, corn, potato, soybean, petunia, tomato,and tobacco can also be utilized as hosts.

In another aspect, expression of humanized anti-BAFF is carried out invertebrate cells. The propagation of vertebrate cells in culture (tissueculture) has become routine procedure and techniques are widelyavailable. Examples of useful mammalian host cell lines are monkeykidney CV1 line transformed by SV40 (COS-7, ATCC CRL 1651), humanembryonic kidney line (293 or 293 cells subcloned for growth insuspension culture, (Graham et al., 1977, J. Gen Virol. 36: 59), babyhamster kidney cells (BHK, ATCC CCL 10), Chinese hamster ovarycells/−DHFR1 (CHO, Urlaub et al., 1980, Proc. Natl. Acad. Sci. USA 77:4216; e.g., DG44), mouse sertoli cells (TM4, Mather, 1980, Biol. Reprod.23:243-251), monkey kidney cells (CV1 ATCC CCL 70), African green monkeykidney cells (VERO-76, ATCC CRL-1587), human cervical carcinoma cells(HELA, ATCC CCL 2), canine kidney cells (MDCK, ATCC CCL 34), buffalo ratliver cells (BRL 3A, ATCC CRL 1442), human lung cells (W138, ATCC CCL75), human liver cells (Hep G2, HB 8065), mouse mammary tumor (MMT060562, ATCC CCL51), TR1 cells (Mather et al., 1982, Annals N.Y. Acad.Sci. 383: 44-68), MRC 5 cells, FS4 cells, and human hepatoma line (HepG2).

Host cells are transformed with the above-described expression orcloning vectors for humanized anti-BAFF antibody production and culturedin conventional nutrient media modified as appropriate for inducingpromoters, selecting transformants, or amplifying the genes encoding thedesired sequences.

The host cells used to produce a humanized anti-BAFF antibody describedherein may be cultured in a variety of media. Commercially availablemedia such as Ham's F10 (Sigma-Aldrich Co., St. Louis, Mo.), MinimalEssential Medium ((MEM), (Sigma-Aldrich Co.), RPMI-1640 (Sigma-AldrichCo.), and Dulbecco's Modified Eagle's Medium ((DMEM), Sigma-Aldrich Co.)are suitable for culturing the host cells. In addition, any of the mediadescribed in one or more of Ham et al., 1979, Meth. Enz. 58: 44, Barneset al., 1980, Anal. Biochem. 102: 255, U.S. Pat. No. 4,767,704, U.S.Pat. No. 4,657,866, U.S. Pat. No. 4,927,762, U.S. Pat. No. 4,560,655,U.S. Pat. No. 5,122,469, WO 90/103430, and WO 87/00195 may be used asculture media for the host cells. Any of these media may be supplementedas necessary with hormones and/or other growth factors (such as insulin,transferrin, or epidermal growth factor), salts (such as sodiumchloride, calcium, magnesium, and phosphate), buffers (such as HEPES),nucleotides (such as adenosine and thymidine), antibiotics (such asgentamicin), trace elements (defined as inorganic compounds usuallypresent at final concentrations in the micromolar range), and glucose oran equivalent energy source. Other supplements may also be included atappropriate concentrations that would be known to those skilled in theart. The culture conditions, such as temperature, pH, and the like, arethose previously used with the host cell selected for expression, andwill be apparent to the ordinarily skilled artisan.

When using recombinant techniques, the antibody can be producedintracellularly, in the periplasmic space, or directly secreted into themedium. If the antibody is produced intracellularly, the cells may bedisrupted to release protein as a first step. Particulate debris, eitherhost cells or lysed fragments, can be removed, for example, bycentrifugation or ultrafiltration. Carter et al., 1992, Bio/Technology10:163-167 describes a procedure for isolating antibodies that aresecreted to the periplasmic space of E. coli. Briefly, cell paste isthawed in the presence of sodium acetate (pH 3.5), EDTA, andphenylmethylsulfonylfluoride (PMSF) over about 30 minutes. Cell debriscan be removed by centrifugation. Where the antibody is secreted intothe medium, supernatants from such expression systems are generallyfirst concentrated using a commercially available protein concentrationfilter, for example, an Amicon or Millipore Pellicon ultrafiltrationunit. A protease inhibitor such as PMSF may be included in any of theforegoing steps to inhibit proteolysis and antibiotics may be includedto prevent the growth of adventitious contaminants. A variety of methodscan be used to isolate the antibody from the host cell.

The antibody composition prepared from the cells can be purified using,for example, hydroxylapatite chromatography, gel electrophoresis,dialysis, and affinity chromatography, with affinity chromatographybeing a typical purification technique. The suitability of protein A asan affinity ligand depends on the species and isotype of anyimmunoglobulin Fc domain that is present in the antibody. Protein A canbe used to purify antibodies that are based on human gamma1, gamma2, orgamma4 heavy chains (see, e.g., Lindmark et al., 1983 J. Immunol. Meth.62:1-13). Protein G is recommended for all mouse isotypes and for humangamma3 (see, e.g., Guss et al., 1986 EMBO J. 5:1567-1575). A matrix towhich an affinity ligand is attached is most often agarose, but othermatrices are available. Mechanically stable matrices such as controlledpore glass or poly(styrenedivinyl)benzene allow for faster flow ratesand shorter processing times than can be achieved with agarose. Wherethe antibody comprises a C_(H3) domain, the Bakerbond ABX™ resin (J. T.Baker, Phillipsburg, N.J.) is useful for purification. Other techniquesfor protein purification such as fractionation on an ion-exchangecolumn, ethanol precipitation, reverse phase HPLC, chromatography onsilica, chromatography on heparin SEPHAROSE™ chromatography on an anionor cation exchange resin (such as a polyaspartic acid column),chromatofocusing, SDS-PAGE, and ammonium sulfate precipitation are alsoavailable depending on the antibody to be recovered.

Following any preliminary purification step(s), the mixture comprisingthe antibody of interest and contaminants may be subjected to low pHhydrophobic interaction chromatography using an elution buffer at a pHbetween about 2.5-4.5, typically performed at low salt concentrations(e.g., from about 0-0.25M salt).

Also included are nucleic acids that hybridize under low, moderate, andhigh stringency conditions, as defined herein, to all or a portion(e.g., the portion encoding the variable region) of the nucleotidesequence represented by isolated polynucleotide sequence(s) that encodean antibody or antibody fragment of the present invention. Thehybridizing portion of the hybridizing nucleic acid is typically atleast 15 (e.g., 20, 25, 30 or 50) nucleotides in length. The hybridizingportion of the hybridizing nucleic acid is at least 80%, e.g., at least90%, at least 95%, or at least 98%, identical to the sequence of aportion or all of a nucleic acid encoding an anti-BAFF polypeptide(e.g., a heavy chain or light chain variable region), or its complement.Hybridizing nucleic acids of the type described herein can be used, forexample, as a cloning probe, a primer, e.g., a PCR primer, or adiagnostic probe.

Some embodiments include isolated polynucleotides including sequencesthat encode an antibody or antibody fragment having the amino acidsequence of any one of 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93,94, 95, 96, 97, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110,111, 112, 113, 114 or 115 and that is at least 80%, at least 90%, atleast 95%, at least 98%, or at least 99% identical to the polynucleotidesequences of SEQ ID NO: 234, 393, 394, 395, 396, 397 or 398.

As used herein, the terms “identical” or “percent identity,” in thecontext of two or more nucleic acids or polypeptide sequences, refer totwo or more sequences or subsequences that are the same or have aspecified percentage of nucleotides or amino acid residues that are thesame, when compared and aligned for maximum correspondence. To determinethe percent identity, the sequences are aligned for optimal comparisonpurposes (e.g., gaps can be introduced in the sequence of a first aminoacid or nucleic acid sequence for optimal alignment with a second aminoor nucleic acid sequence). The amino acid residues or nucleotides atcorresponding amino acid positions or nucleotide positions are thencompared. When a position in the first sequence is occupied by the sameamino acid residue or nucleotide as the corresponding position in thesecond sequence, then the molecules are identical at that position. Thepercent identity between the two sequences is a function of the numberof identical positions shared by the sequences (i.e., % identity=# ofidentical positions/total # of positions (e.g., overlappingpositions)×100). In some embodiments, the two sequences that arecompared are the same length after gaps are introduced within thesequences, as appropriate (e.g., excluding additional sequence extendingbeyond the sequences being compared). For example, when variable regionsequences are compared, the leader and/or constant domain sequences arenot considered. For sequence comparisons between two sequences, a“corresponding” CDR refers to a CDR in the same location in bothsequences (e.g., CDR-H1 of each sequence).

The determination of percent identity or percent similarity between twosequences can be accomplished using a mathematical algorithm. Apreferred, non-limiting example of a mathematical algorithm utilized forthe comparison of two sequences is the algorithm of Karlin and Altschul,1990, Proc. Natl. Acad. Sci. USA 87:2264-2268, modified as in Karlin andAltschul, 1993, Proc. Natl. Acad. Sci. USA 90:5873-5877. Such analgorithm is incorporated into the NBLAST and XBLAST programs ofAltschul et al., 1990, J. Mol. Biol. 215:403-410. BLAST nucleotidesearches can be performed with the NBLAST program, score=100,wordlength=12, to obtain nucleotide sequences homologous to a nucleicacid encoding a protein of interest. BLAST protein searches can beperformed with the XBLAST program, score=50, wordlength=3, to obtainamino acid sequences homologous to protein of interest. To obtain gappedalignments for comparison purposes, Gapped BLAST can be utilized asdescribed in Altschul et al., 1997, Nucleic Acids Res. 25:3389-3402.Alternatively, PSI-Blast can be used to perform an iterated search whichdetects distant relationships between molecules (Id.). When utilizingBLAST, Gapped BLAST, and PSI-Blast programs, the default parameters ofthe respective programs (e.g., XBLAST and NBLAST) can be used. Anotherpreferred, non-limiting example of a mathematical algorithm utilized forthe comparison of sequences is the algorithm of Myers and Miller, CABIOS(1989). Such an algorithm is incorporated into the ALIGN program(version 2.0) which is part of the GCG sequence alignment softwarepackage. When utilizing the ALIGN program for comparing amino acidsequences, a PAM120 weight residue table, a gap length penalty of 12,and a gap penalty of 4 can be used. Additional algorithms for sequenceanalysis are known in the art and include ADVANCE and ADAM as describedin Torellis and Robotti, 1994, Comput. Appl. Biosci. 10:3-5; and FASTAdescribed in Pearson and Lipman, 1988, Proc. Natl. Acad. Sci. USA85:2444-8. Within FASTA, ktup is a control option that sets thesensitivity and speed of the search. If ktup=2, similar regions in thetwo sequences being compared are found by looking at pairs of alignedresidues; if ktup=1, single aligned amino acids are examined. ktup canbe set to 2 or 1 for protein sequences, or from 1 to 6 for DNAsequences. The default if ktup is not specified is 2 for proteins and 6for DNA. Alternatively, protein sequence alignment may be carried outusing the CLUSTAL W algorithm, as described by Higgins et al., 1996,Methods Enzymol. 266:383-402.

Non-Therapeutic Uses

The antibodies described herein are useful as affinity purificationagents. In this process, the antibodies are immobilized on a solid phasesuch a Protein A resin, using methods well known in the art. Theimmobilized antibody is contacted with a sample containing the BAFFprotein (or fragment thereof) to be purified, and thereafter the supportis washed with a suitable solvent that will remove substantially all thematerial in the sample except the BAFF protein, which is bound to theimmobilized antibody. Finally, the support is washed with anothersuitable solvent that will release the BAFF protein from the antibody.

Anti-BAFF antibodies, for example humanized anti-BAFF antibodies, arealso useful in diagnostic assays to detect and/or quantify BAFF protein,for example, detecting BAFF expression in specific cells, tissues, orserum. The anti-BAFF antibodies can be used diagnostically to, forexample, monitor the development or progression of a disease as part ofa clinical testing procedure to, e.g., determine the efficacy of a giventreatment and/or prevention regimen. Detection can be facilitated bycoupling the anti-BAFF antibody. Examples of detectable substancesinclude various enzymes, prosthetic groups, fluorescent materials,luminescent materials, bioluminescent materials, radioactive materials,positron emitting metals using various positron emission tomographies,and nonradioactive paramagnetic metal ions. See, for example, U.S. Pat.No. 4,741,900 for metal ions which can be conjugated to antibodies foruse as diagnostics according to the present invention.

The anti-BAFF antibodies can be used in methods for diagnosing aBAFF-associated disorder (e.g., a disorder characterized by abnormalexpression of BAFF) or to determine if a subject has an increased riskof developing a BAFF-associated disorder. Such methods includecontacting a biological sample from a subject with a BAFF antibody anddetecting binding of the antibody to BAFF. By “biological sample” isintended any biological sample obtained from an individual, cell line,tissue culture, or other source of cells potentially expressing BAFF.Methods for obtaining tissue biopsies and body fluids from mammals arewell known in the art.

In some embodiments, the method can further comprise comparing the levelof BAFF in a patient sample to a control sample (e.g., a subject thatdoes not have a BAFF-associated disorder) to determine if the patienthas a BAFF-associated disorder or is at risk of developing aBAFF-associated disorder.

It will be advantageous in some embodiments, for example, for diagnosticpurposes to label the antibody with a detectable moiety. Numerousdetectable labels are available, including radioisotopes, fluorescentlabels, enzyme substrate labels and the like. The label may beindirectly conjugated with the antibody using various known techniques.For example, the antibody can be conjugated with biotin and any of thethree broad categories of labels mentioned above can be conjugated withavidin, or vice versa. Biotin binds selectively to avidin and thus, thelabel can be conjugated with the antibody in this indirect manner.Alternatively, to achieve indirect conjugation of the label with theantibody, the antibody can be conjugated with a small hapten (such asdigoxin) and one of the different types of labels mentioned above isconjugated with an anti-hapten antibody (e.g., anti-digoxin antibody).Thus, indirect conjugation of the label with the antibody can beachieved.

Exemplary radioisotopes labels include ³⁵S, ¹⁴C, ¹²⁵I, ³H, and ¹³¹I. Theantibody can be labeled with the radioisotope, using the techniquesdescribed in, for example, Current Protocols in Immunology, Volumes 1and 2, 1991, Coligen et al., Ed. Wiley-Interscience, New York, N.Y.,Pubs. Radioactivity can be measured, for example, by scintillationcounting.

Exemplary fluorescent labels include labels derived from rare earthchelates (europium chelates) or fluorescein and its derivatives,rhodamine and its derivatives, dansyl, Lissamine, phycoerythrin, andTexas Red are available. The fluorescent labels can be conjugated to theantibody via known techniques, such as those disclosed in CurrentProtocols in Immunology, for example. Fluorescence can be quantifiedusing a fluorimeter.

There are various well-characterized enzyme-substrate labels known inthe art (see, e.g., U.S. Pat. No. 4,275,149). The enzyme generallycatalyzes a chemical alteration of the chromogenic substrate that can bemeasured using various techniques. For example, alteration may be acolor change in a substrate that can be measured spectrophotometrically.Alternatively, the enzyme may alter the fluorescence orchemiluminescence of the substrate. Techniques for quantifying a changein fluorescence are described above. The chemiluminescent substratebecomes electronically excited by a chemical reaction and may then emitlight that can be measured, using a chemiluminometer, for example, ordonates energy to a fluorescent acceptor.

Examples of enzymatic labels include luciferases such as fireflyluciferase and bacterial luciferase (U.S. Pat. No. 4,737,456),luciferin, 2,3-dihydrophthalazinediones, malate dehydrogenase, urease,peroxidase such as horseradish peroxidase (HRPO), alkaline phosphatase,β-galactosidase, glucoamylase, lysozyme, saccharide oxidases (such asglucose oxidase, galactose oxidase, and glucose-6-phosphatedehydrogenase), heterocydic oxidases (such as uricase and xanthineoxidase), lactoperoxidase, microperoxidase, and the like. Techniques forconjugating enzymes to antibodies are described, for example, inO'Sullivan et al., 1981, Methods for the Preparation of Enzyme-AntibodyConjugates for use in Enzyme Immunoassay, in Methods in Enzym. (J.Langone & H. Van Vunakis, eds.), Academic press, N.Y., 73: 147-166.

Examples of enzyme-substrate combinations include, for example:Horseradish peroxidase (HRPO) with hydrogen peroxidase as a substrate,wherein the hydrogen peroxidase oxidizes a dye precursor such asorthophenylene diamine (OPD) or 3,3′,5,5′-tetramethyl benzidinehydrochloride (TMB); alkaline phosphatase (AP) with para-Nitrophenylphosphate as chromogenic substrate; and β-D-galactosidase (β-D-Gal) witha chromogenic substrate such as p-nitrophenyl-β-D-galactosidase orfluorogenic substrate 4-methylumbelliferyl-β-D-galactosidase.

Numerous other enzyme-substrate combinations are available to thoseskilled in the art. For a general review of these, see U.S. Pat. No.4,275,149 and U.S. Pat. No. 4,318,980.

In another embodiment, the humanized anti-BAFF antibody is usedunlabeled and detected with a labeled antibody that binds the humanizedanti-BAFF antibody.

The antibodies described herein may be employed in any known assaymethod, such as competitive binding assays, direct and indirect sandwichassays, and immunoprecipitation assays. See, e.g., Zola, MonoclonalAntibodies: A Manual of Techniques, pp. 147-158 (CRC Press, Inc. 1987).

The anti-BAFF antibody or antigen binding fragment thereof can be usedto inhibit the binding of BAFF to one of the BAFF receptors. Suchmethods comprise administering an anti-BAFF antibody or antigen bindingfragment thereof to a cell (e.g., a mammalian cell) or cellularenvironment, whereby signaling mediated by the BAFF receptor isinhibited. These methods can be performed in vitro or in vivo. By“cellular environment” is intended the tissue, medium, or extracellularmatrix surrounding a cell. The anti-BAFF antibody or antigen bindingfragment thereof is administered to the cellular environment of a cellin such a manner that the antibody or fragment is capable of binding toBAFF molecules outside of and surrounding the cell, therefore,preventing the binding of BAFF to its receptor.

Diagnostic Kits

An anti-BAFF antibody can be used in a diagnostic kit, i.e., a packagedcombination of reagents in predetermined amounts with instructions forperforming the diagnostic assay. Where the antibody is labeled with anenzyme, the kit may include substrates and cofactors required by theenzyme such as a substrate precursor that provides the detectablechromophore or fluorophore. In addition, other additives may be includedsuch as stabilizers, buffers (for example a block buffer or lysisbuffer), and the like. The relative amounts of the various reagents maybe varied widely to provide for concentrations in solution of thereagents that substantially optimize the sensitivity of the assay. Thereagents may be provided as dry powders, usually lyophilized, includingexcipients that on dissolution will provide a reagent solution havingthe appropriate concentration.

Therapeutic Uses

In another embodiment, a humanized anti-BAFF antibody disclosed hereinis useful in the treatment of various disorders associated with theexpression of BAFF as described herein. Methods for treating a BAFFassociated disorder comprise administering a therapeutically effectiveamount of a humanized anti-BAFF antibody to a subject in need thereof.

The humanized anti-BAFF antibody or agent is administered by anysuitable means, including parenteral, subcutaneous, intraperitoneal,intrapulmonary, and intranasal, and, if desired for localimmunosuppressive treatment, intralesional administration (includingperfusing or otherwise contacting the graft with the antibody beforetransplantation). The humanized anti-BAFF antibody or agent can beadministered, for example, as an infusion or as a bolus. Parenteralinfusions include intramuscular, intravenous, intraarterial,intraperitoneal, or subcutaneous administration. In addition, thehumanized anti-BAFF antibody is suitably administered by pulse infusion,particularly with declining doses of the antibody. In one aspect, thedosing is given by injections, most preferably intravenous orsubcutaneous injections, depending in part on whether the administrationis brief or chronic.

For the prevention or treatment of disease, the appropriate dosage ofantibody will depend on a variety of factors such as the type of diseaseto be treated, as defined above, the severity and course of the disease,whether the antibody is administered for preventive or therapeuticpurposes, previous therapy, the patient's clinical history and responseto the antibody, and the discretion of the attending physician. Theantibody is suitably administered to the patient at one time or over aseries of treatments.

Depending on the type and severity of the disease, about 1 μg/kg to 20mg/kg (e.g., 0.1-15 mg/kg) of antibody is an initial candidate dosagefor administration to the patient, whether, for example, by one or moreseparate administrations, or by continuous infusion. A typical dailydosage might range from about 1 μg/kg to 100 mg/kg or more, depending onthe factors mentioned above. For repeated administrations over severaldays or longer, depending on the condition, the treatment is sustaineduntil a desired suppression of disease symptoms occurs. However, otherdosage regimens may be useful. The progress of this therapy is easilymonitored by conventional techniques and assays. An exemplary dosingregimen is that disclosed in WO 94/04188.

The term “suppression” is used herein in the same context as“amelioration” and “alleviation” to mean a lessening of one or morecharacteristics of the disease.

The antibody composition will be formulated, dosed, and administered ina fashion consistent with good medical practice. Factors forconsideration in this context include the particular disorder beingtreated, the particular mammal being treated, the clinical condition ofthe individual patient, the cause of the disorder, the site of deliveryof the agent, the method of administration, the scheduling ofadministration, and other factors known to medical practitioners. The“therapeutically effective amount” of the antibody to be administeredwill be governed by such considerations, and is the minimum amountnecessary to prevent, ameliorate, or treat the disorder associated withBAFF expression.

The antibody need not be, but is optionally, formulated with one or moreagents currently used to prevent or treat the disorder in question. Theeffective amount of such other agents depends on the amount of humanizedanti-BAFF antibody present in the formulation, the type of disorder ortreatment, and other factors discussed above. These are generally usedin the same dosages and with administration routes as used hereinbeforeor about from 1 to 99% of the heretofore employed dosages.

BAFF-Associated Disorders

The anti-BAFF antibodies or agents are useful for treating or preventingan immunological disorder characterized by abnormal expression of BAFF.The anti-BAFF antibodies or antigen binding fragments thereof also finduse in the treatment or prevention of respiratory disorders, metabolicdisorders, for example diabetes mellitus, and certain cancers. Treatmentor prevention of the immunological disorder, respiratory disorder,metabolic disorder or cancer, according to the methods described herein,is achieved by administering to a subject in need of such treatment orprevention an effective amount of the anti-BAFF antibody or agent,whereby the antibody decreases the activity of BAFF associated with thedisease state.

Immunological diseases that are characterized by inappropriateactivation of immune cells and that can be treated or prevented by themethods described herein can be classified, for example, by the type(s)of hypersensitivity reaction(s) that underlie the disorder. Thesereactions are typically classified into four types: anaphylacticreactions, cytotoxic (cytolytic) reactions, immune complex reactions, orcell-mediated immunity (CMI) reactions (also referred to as delayed-typehypersensitivity (DTH) reactions). (See, e.g., Fundamental Immunology(William E. Paul ed., Raven Press, N.Y., 3rd ed. 1993).) Immunologicaldiseases include inflammatory diseases and autoimmune diseases.

Specific examples of such immunological diseases include the following:rheumatoid arthritis, autoimmune demyelinative diseases (e.g., multiplesclerosis, allergic encephalomyelitis), endocrine opthalmopathy,uveoretinitis, systemic lupus erythematosus, myasthenia gravis, Grave'sdisease, glomerulonephritis, autoimmune hepatological disorder,inflammatory bowel disease (e.g., Crohn's disease or ulcerativecolitis), anaphylaxis, allergic reaction, Sjogren's syndrome, type Idiabetes mellitus, primary biliary cirrhosis, Wegener's granulomatosis,fibromyalgia, polymyositis, dermatomyositis, inflammatory myositis,multiple endocrine failure, Schmidt's syndrome, autoimmune uveitis,Addison's disease, adrenalitis, thyroiditis, Hashimoto's thyroiditis,autoimmune thyroid disease, pernicious anemia, gastric atrophy, chronichepatitis, lupoid hepatitis, atherosclerosis, subacute cutaneous lupuserythematosus, hypoparathyroidism, Dressler's syndrome, autoimmunethrombocytopenia, idiopathic thrombocytopenic purpura, hemolytic anemia,pemphigus vulgaris, pemphigus, dermatitis herpetiformis, alopeciaarcata, pemphigoid, scleroderma, progressive systemic sclerosis, CRESTsyndrome (calcinosis, Raynaud's phenomenon, esophageal dysmotility,sclerodactyl), and telangiectasia), male and female autoimmuneinfertility, ankylosing spondolytis, ulcerative colitis, mixedconnective tissue disease, polyarteritis nedosa, systemic necrotizingvasculitis, atopic dermatitis, atopic rhinitis, Goodpasture's syndrome,Chagas' disease, sarcoidosis, rheumatic fever, asthma, recurrentabortion, anti-phospholipid syndrome, farmer's lung, erythemamultiforme, post cardiotomy syndrome, Cushing's syndrome, autoimmunechronic active hepatitis, bird-fancier's lung, toxic epidermalnecrolysis, Alport's syndrome, alveolitis, allergic alveolitis,fibrosing alveolitis, interstitial lung disease, erythema nodosum,pyoderma gangrenosum, transfusion reaction, Takayasu's arteritis,polymyalgia rheumatica, temporal arteritis, schistosomiasis, giant cellarteritis, ascariasis, aspergillosis, Sampter's syndrome, eczema,lymphomatoid granulomatosis, Behcet's disease, Caplan's syndrome,Kawasaki's disease, dengue, encephalomyelitis, endocarditis,endomyocardial fibrosis, endophthalmitis, erythema elevatum et diutinum,psoriasis, psoriatic arthritis, erythroblastosis fetalis, eosinophilicfaciitis, Shulman's syndrome, Felty's syndrome, filariasis, cyclitis,chronic cyclitis, heterochronic cyclitis, Fuch's cyclitis, IgAnephropathy, Henoch-Schonlein purpura, graft versus host disease,anti-neutrophil cytoplasmic antibodies (ANCA)-associated vasculitis,transplantation rejection, cardiomyopathy, Eaton-Lambert syndrome,relapsing polychondritis, cryoglobulinemia, Waldenstrom'smacroglobulemia, Evan's syndrome, acute respiratory distress syndrome,pulmonary inflammation, osteoporosis, delayed type hypersensitivity andautoimmune gonadal failure.

In another aspect, the anti-BAFF antibodies and agents as describedherein are also useful for treating cancers, in which BAFF is abnormallyexpressed.

BAFF-expressing cancers that can be treated by the methods describedherein include, for example, leukemia, such as acute leukemia, acutelymphocytic leukemia, acute myelocytic leukemia (e.g., myeloblastic,promyelocytic, myelomonocytic, monocytic, or erythroleukemia), chronicleukemia, chronic myelocytic (granulocytic) leukemia, or chroniclymphocytic leukemia; Polycythemia vera; Lymphoma (e.g., Hodgkin'sdisease or Non-Hodgkin's disease); multiple myeloma, Waldenstrom'smacroglobulinemia; heavy chain disease; solid tumors such sarcomas andcarcinomas (e.g., fibrosarcoma, myxosarcoma, liposarcoma,chondrosarcoma, osteogenic sarcoma, osteosarcoma, chordoma,angiosarcoma, endotheliosarcoma, lymphangiosarcoma,lymphangioendotheliosarcoma, synovioma, mesothelioma, Ewing's tumor,leiomyosarcoma, rhabdomyosarcoma, colon carcinoma, colorectal carcinoma,pancreatic cancer, breast cancer, ovarian cancer, prostate cancer,squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweatgland carcinoma, sebaceous gland carcinoma, papillary carcinoma,papillary adenocarcinomas, cystadenocarcinoma, medullary carcinoma,bronchogenic carcinoma, renal cell carcinoma, hepatoma, bile ductcarcinoma, choriocarcinoma, seminoma, embryonal carcinoma, Wilms' tumor,cervical cancer, uterine cancer, testicular tumor, lung carcinoma, smallcell lung carcinoma, non-small cell lung carcinoma, bladder carcinoma,epithelial carcinoma, glioma, astrocytoma, medulloblastoma,craniopharyngioma, ependymoma, pinealoma, hemangioblastoma, acousticneuroma, oligodendroglioma, menangioma, melanoma, neuroblastoma,retinoblastoma, nasopharyngeal carcinoma, or esophageal carcinoma).

Pharmaceutical Compositions and Administration Thereof

A composition comprising a BAFF binding agent (e.g., an anti-BAFFantibody) can be administered to a subject having or at risk of havingan immunological disorder, respiratory disorder or a cancer. Theinvention further provides for the use of a BAFF binding agent (e.g., ananti-BAFF antibody) in the manufacture of a medicament for prevention ortreatment of a cancer, respiratory disorder or immunological disorder.The term “subject” as used herein means any mammalian patient to which aBAFF binding agent can be administered, including, e.g., humans andnon-human mammals, such as primates, rodents, and dogs. Subjectsspecifically intended for treatment using the methods described hereininclude humans. The antibodies or agents can be administered eitheralone or in combination with other compositions in the prevention ortreatment of the immunological disorder, respiratory disorder or cancer.Such compositions which can be administered in combination with theantibodies or agents include methotrexate (MTX) and immunomodulators,e.g. antibodies or small molecules.

Examples of antibodies for use in such pharmaceutical compositions arethose that comprise a humanized antibody or antibody fragment having thelight chain variable region amino acid sequence of any of SEQ ID NO: 82,83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, or 97. Examplesof antibodies for use in such pharmaceutical compositions are also thosethat comprise a humanized antibody or antibody fragment having the heavychain variable region amino acid sequence of any of SEQ ID NO: 100, 101,102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114 or 115.

Various delivery systems are known and can be used to administer theBAFF binding agent. Methods of introduction include but are not limitedto intradermal, intramuscular, intraperitoneal, intravenous,subcutaneous, intranasal, epidural, and oral routes. The BAFF bindingagent can be administered, for example by infusion, bolus or injection,and can be administered together with other biologically active agentssuch as chemotherapeutic agents. Administration can be systemic orlocal. In preferred embodiments, the administration is by subcutaneousinjection. Formulations for such injections may be prepared in forexample prefilled syringes that may be administered once every otherweek.

In specific embodiments, the BAFF binding agent composition isadministered by injection, by means of a catheter, by means of asuppository, or by means of an implant, the implant being of a porous,non-porous, or gelatinous material, including a membrane, such as asialastic membrane, or a fiber. Typically, when administering thecomposition, materials to which the anti-BAFF antibody or agent does notabsorb are used.

In other embodiments, the anti-BAFF antibody or agent is delivered in acontrolled release system. In one embodiment, a pump may be used (see,e.g., Langer, 1990, Science 249:1527-1533; Sefton, 1989, CRC Crit. Ref.Biomed. Eng. 14:201; Buchwald et al., 1980, Surgery 88:507; Saudek etal., 1989, N. Engl. J. Med. 321:574). In another embodiment, polymericmaterials can be used. (See, e.g., Medical Applications of ControlledRelease (Langer and Wise eds., CRC Press, Boca Raton, Fla., 1974);Controlled Drug Bioavailability, Drug Product Design and Performance(Smolen and Ball eds., Wiley, New York, 1984); Ranger and Peppas, 1983,Macromol. Sci. Rev. Macromol. Chem. 23:61. See also Levy et al., 1985,Science 228:190; During et al., 1989, Ann. Neurol. 25:351; Howard etal., 1989, J. Neurosurg. 71:105.) Other controlled release systems arediscussed, for example, in Langer, supra.

An BAFF binding agent (e.g., an anti-BAFF antibody) can be administeredas pharmaceutical compositions comprising a therapeutically effectiveamount of the binding agent and one or more pharmaceutically compatibleingredients.

In typical embodiments, the pharmaceutical composition is formulated inaccordance with routine procedures as a pharmaceutical compositionadapted for intravenous or subcutaneous administration to human beings.Typically, compositions for administration by injection are solutions insterile isotonic aqueous buffer. Where necessary, the pharmaceutical canalso include a solubilizing agent and a local anesthetic such aslignocaine to ease pain at the site of the injection. Generally, theingredients are supplied either separately or mixed together in unitdosage form, for example, as a dry lyophilized powder or water freeconcentrate in a hermetically sealed container such as an ampoule orsachette indicating the quantity of active agent. Where thepharmaceutical is to be administered by infusion, it can be dispensedwith an infusion bottle containing sterile pharmaceutical grade water orsaline. Where the pharmaceutical is administered by injection, anampoule of sterile water for injection or saline can be provided so thatthe ingredients can be mixed prior to administration.

Further, the pharmaceutical composition can be provided as apharmaceutical kit comprising (a) a container containing a BAFF bindingagent (e.g., an anti-BAFF antibody) in lyophilized form and (b) a secondcontainer containing a pharmaceutically acceptable diluent (e.g.,sterile water) for injection. The pharmaceutically acceptable diluentcan be used for reconstitution or dilution of the lyophilized anti-BAFFantibody or agent. Optionally associated with such container(s) can be anotice in the form prescribed by a governmental agency regulating themanufacture, use or sale of pharmaceuticals or biological products,which notice reflects approval by the agency of manufacture, use or salefor human administration.

The amount of the BAFF binding agent (e.g., anti-BAFF antibody) that iseffective in the treatment or prevention of an immunological disorder orcancer can be determined by standard clinical techniques. In addition,in vitro assays may optionally be employed to help identify optimaldosage ranges. The precise dose to be employed in the formulation willalso depend on the route of administration, and the stage ofimmunological disorder or cancer, and should be decided according to thejudgment of the practitioner and each patient's circumstances. Effectivedoses may be extrapolated from dose-response curves derived from invitro or animal model test systems.

Generally, the dosage of an anti-BAFF antibody or BAFF binding agentadministered to a patient with an immunological disorder is typicallyabout 0.1 mg/kg to about 100 mg/kg of the subject's body weight. Thedosage administered to a subject is about 0.1 mg/kg to about 50 mg/kg,about 1 mg/kg to about 30 mg/kg, about 1 mg/kg to about 20 mg/kg, about1 mg/kg to about 15 mg/kg, or about 1 mg/kg to about 10 mg/kg of thesubject's body weight.

Exemplary doses include, but are not limited to, from 1 ng/kg to 100mg/kg. In some embodiments, a dose is about 0.5 mg/kg, about 1 mg/kg,about 2 mg/kg, about 3 mg/kg, about 4 mg/kg, about 5 mg/kg, about 6mg/kg, about 7 mg/kg, about 8 mg/kg, about 9 mg/kg, about 10 mg/kg,about 11 mg/kg, about 12 mg/kg, about 13 mg/kg, about 14 mg/kg, about 15mg/kg or about 16 mg/kg. The dose can be administered, for example,daily, once per week (weekly), twice per week, thrice per week, fourtimes per week, five times per week, six times per week, biweekly ormonthly, every two months, or every three months. In specificembodiments, the dose is about 0.5 mg/kg/week, about 1 mg/kg/week, about2 mg/kg/week, about 3 mg/kg/week, about 4 mg/kg/week, about 5mg/kg/week, about 6 mg/kg/week, about 7 mg/kg/week, about 8 mg/kg/week,about 9 mg/kg/week, about 10 mg/kg/week, about 11 mg/kg/week, about 12mg/kg/week, about 13 mg/kg/week, about 14 mg/kg/week, about 15mg/kg/week or about 16 mg/kg/week. In some embodiments, the dose rangesfrom about 1 mg/kg/week to about 15 mg/kg/week.

In some embodiments, the pharmaceutical compositions comprising the BAFFbinding agent can further comprise a therapeutic agent, eitherconjugated or unconjugated to the binding agent. The anti-BAFF antibodyor BAFF binding agent can be co-administered in combination with one ormore therapeutic agents for the treatment or prevention of immunologicaldisorders or cancers.

Such combination therapy administration can have an additive orsynergistic effect on disease parameters (e.g., severity of a symptom,the number of symptoms, or frequency of relapse). With respect totherapeutic regimens for combinatorial administration, in a specificembodiment, an anti-BAFF antibody or BAFF binding agent is administeredconcurrently with a therapeutic agent. In another specific embodiment,the therapeutic agent is administered prior or subsequent toadministration of the anti-BAFF antibody or BAFF binding agent, by atleast an hour and up to several months, for example at least an hour,five hours, 12 hours, a day, a week, a month, or three months, prior orsubsequent to administration of the anti-BAFF antibody or BAFF bindingagent.

Articles of Manufacture

In another aspect, an article of manufacture containing materials usefulfor the treatment of the disorders described above is included. Thearticle of manufacture comprises a container and a label. Suitablecontainers include, for example, bottles, vials, syringes, and testtubes. The containers may be formed from a variety of materials such asglass or plastic. The container holds a composition that is effectivefor treating the condition and may have a sterile access port. Forexample, the container may be an intravenous solution bag or a vialhaving a stopper pierceable by a hypodermic injection needle. The activeagent in the composition is the humanized anti-BAFF antibody. The labelon or associated with the container indicates that the composition isused for treating the condition of choice. The article of manufacturemay further comprise a second container comprising apharmaceutically-acceptable buffer, such as phosphate-buffered saline,Ringer's solution, and dextrose solution. It may further include othermaterials desirable from a commercial and user standpoint, includingother buffers, diluents, filters, needles, syringes, and package insertswith instructions for use.

The invention is further described in the following examples, which arenot intended to limit the scope of the invention.

EXAMPLES Example 1 Generation of Mouse Antibodies

The lead mouse antibodies of the present invention were derived frommouse hybridomas. Various strains of mice were immunized multiple timesfor up to 6 months. Immunization of mice was carried out using suitabletechniques contained in the art. For example, to obtain a specificimmunogenic response, various versions of recombinant soluble human BAFFprotein (amino acids 72-285) were used, including human BAFF fusionprotein antigen immunizations. In addition, some mice were immunizedwith a mouse cell line transfected to express human BAFF on the cellsurface. Preparation of immunogenic antigens including adjuvants andimmunization routes can also be performed using suitable techniquesknown in the art. Serum binding titers of sufficient requirements weremet, and mouse lymphocytes were fused to mouse myeloma cells usingvarious methods of the established art. Screening of hybridomas wasperformed to obtained high affinity, and specific antibodies.

Example 2 Generation of Humanized Anti-BAFF Fabs

Mouse lead antibodies 13J018 and 235F5 were converted to a chimericantibody consisting of the mouse variable domains of 1A4 and 5B9,respectively, and a human constant IgG1KO domain. Mouse antibodies 1A4and 5B9 are shown in Tables 3 and 4 above. The IgG1KO (knock out) hastwo replacement mutations (Leu234Ala and Leu235A1a) that eliminate ADCCand CDC activity by reducing effector functions such as FcγR andcomplement binding. The variable domains of the mouse and chimericantibodies are identical. Chimeric antibodies are generated to confirmthe function of the antibody and to ensure the correct sequences havebeen obtained. Once correct sequences were identified, the mousevariable domains were used to generate chimeric Fab wherein mouse Vk andVh residues were in frame with human Ck and Ch1 residues respectively.These chimeric Fabs were used as benchmark molecules to screen thehumanized Fabs during the screening process. Next, the mouse variableregions (Vk and Vh) were then humanized through a design and screeningprocess. A library was made where human and mouse residues were variedin such a way that in any given position there could be either a humanor mouse residue. Such a library was made for those amino acids thatwere different between human germline and mouse antibody. Only theclones that retain the function of the parent mouse antibody wereselected using the chimeric Fab. Representative humanized variableregions for antibodies 1A4(13 J018) and 5B9(235F5) are shown in Tables 5and 6.

Example 3 Generation of Recombinant Soluble Trimeric Human BAFF Protein

Human BAFF(72-285) with an N-terminal His-tag (SEQ ID: 398) wasexpressed transiently in HEK293-6E cells through standard lipid-basedtransfection. 96-hours post transfection, cells were pelleted andexpression of protein in the supernatant was verified via an anti 6×Hiswestern blot (“6×His” disclosed as SEQ ID NO: 407). Supernatantpurification was completed using Ni-Sepharose Affinity Chromatography.Purified His-BAFF was cleaved with His-tagged furin protease to produceC-terminal fragment (amino acids 134-285. SEQ ID: 399). To remove furinand cleaved N-terminal fragment species from the sample, the totalprotein sample was passed through an Ni/NTA drip column and the flowthrough was collected. The furin cleaved-huBAFF was polished by SizeExclusion Chromatography. Trimeric status was confirmed by AnalyticalUltracentrifuge analysis.

Sequence for His-tagged human BAFF (72-285): (SEQ ID NO: 399)HHHHHHENLYFQGLQGDLASLRAELQGHHAEKLPAGAGAPKAGLEEAPAVTAGLKIFEPPAPGEGNSSQNSRNKRAVQGPEETVTQDCLQLIADSETPTIQKGSYTFVPWLLSFKRGSALEEKENKILVKETGYFFIYGQVLYTDKTYAMGHLIQRKKVHVFGDELSLVTLFRCIQNMPETLPNNSCYSAGIAKLEEGDELQLAIPRENAQISLDGDVTFFGALKLL Sequence for furin-cleaved human BAFF(134-285): (SEQ ID NO: 400)AVQGPEETVTQDCLQLIADSETPTIQKGSYTFVPWLLSFKRGSALEEKENKILVKETGYFFIYGVLYTDKTYAMGHLIQRKKVHVFGDELSLVTLFRCIQNMPETLPNNSCYSAGIAKLEEGDELQLAIPRENAQISLDGDVTFFGALKL L

Example 4 Binding and Affinity Data for Anti-BAFF Antibodies (Refers toTables 7 and 8)

Apparent binding affinities were evaluated using surface plasmonresonance where the antibodies were captured at different surfacedensities on a Protein A/G surface. Soluble trimeric BAFF at differentconcentrations were flowed over the captured antibody. The kineticvalues are obtained from a global fit of all surface densities using a1:1 Langmuir model and reported in Tables 7 and 8. Clinical referenceantibodies (Reference 1 comprising SEQ ID NOS: 98 and 116) and Reference2 comprising SEQ ID NOS: 99 and 117) were used as comparators.

TABLE 7 Functional Inhibition and Affinity Determination of Anti-BAFFAntibodies. Soluble trimeric Soluble 60-mer huBAFF (52 pM) huBAFF (4.2pM) mbBAFF Neutralization Neutralization Neutralization ApparentAffinity Designation IC90 (pM) n = 2 IC90 (pM) n = 2 IC90 (pM) n = 1K_(D) (pM)** Reference 1 290.0 21.0 1052 <10 Reference 2 1000.0 93% @ 67nM 151000  22.2 206G9A10 35.1 0.2 ND* <10 227D5A7 56.8 0.8 ND <10250E5A11 97.5 0.3 ND <10 235F5B9 107.4 2.2 1050 <10 227D3B11 127.7 1.6ND <10 217H12A7 129.4 13.4 ND <10 210D9B8 155.6 14.0 ND <10 214G4B7296.1 3.0 ND <10 13J018-1A4 304.8 23.0 4650 <10 218H1C10.1 370 41 ND ND218H1C10.2 370 ND ND ND *ND: not determined **at detection limit

TABLE 8 Functional Inhibition and Affinity Determination of Anti-BAFFAntibodies. Soluble trimeric huBAFF (50 pM) Soluble 60-mer huBAFF PotentNeutralization (4 pM) Neutralization mbBAFF Neutralization DesignationIC90 (pM) IC50 (pM) IC90 (pM) IC50 (pM) IC90 (pM) IC50 (pM) Reference 1290 102 40 1052 177 Reference 2 1000 197985 38298 151000 2180 1002E8A6824 107 79 29 2087 141 1070A6B7 677 43 56 20 1065 147 1094C4E6 1099 38420925 4048 175030 2561 27121-3C7 326 55 33 11  ND* ND 317H2A6 327 97 134 ND ND 319B8A12 331 91 24 8 ND ND 320F9C5 3107 110 24 10 ND ND 323E9D1312 148 23 6 ND ND 332C1B12 457 99 40 12 ND ND 344B9D9 352 102 32 11 NDND 348A6C1 329 110 29 9 ND ND 352G11A10 444 99 90 21 ND ND 363D4A10 47321 23 9 ND ND 381A6A9 240 44 42 17 ND ND 384D5A2 765 42 44 15 ND ND394F5A5 433 16 29 7 ND ND 409F12A11 390 31 44 16 ND ND 418F6D9 491 38154 27 ND ND 431G5A3 336 28 100 21 ND ND 435A6B3 294 23 26 10 ND ND436H2C12 408 27 37 15 ND ND 436H6A9 266 26 27 10 ND ND 440E9D12 259 3131 11 ND ND 441E6F2 293 43 37 16 ND ND 443C11A12 309 33 30 13 ND ND444G1A10 284 25 35 12 ND ND 450A2A7 602 36 130 24 ND ND 456H11B7 299 3237 12 ND ND 537G7A6 329 28 39 13 ND ND 551H4D6 257 110 1979 234 ND ND560H2A7 324 35 30 13 ND ND 606H7F8 378 29 44 12 ND ND *ND: notdetermined

Example 5 Functional Inhibition of Antibodies to Soluble Trimeric HumanBAFF (Refers to Tables 7 and 8, and FIG. 1)

Antibodies were assessed for the ability to neutralize soluble trimerichuman BAFF activation of the human BAFF receptor (BAFFR). A fixedconcentration of the trimeric (52 pM) BAFF was mixed in assay mediumwith CHO cells expressing recombinant human BAFFR and a luciferasereporter system and stimulated for 24 hours in an incubator at 37° C.,5% CO2 in the presence of varying doses of anti-BAFF antibodies.Luciferase expression was assessed at the end of the incubation toquantify the level of neutralization achieved. IC50 and IC90 values weredetermined from the plots of the antibody dose titration luciferaseinhibition results. Clinical reference antibodies (Reference 1 andReference 2) were used as comparators.

Example 6 Generation of Recombinant Soluble 60-Mer Human BAFF Protein

Stable HEK293F cells expressing human BAFF(134-285) with an N-terminalHis-tag (SEQ ID: 401) was produced using lentivirus based technologiesfrom Clontech (pLVX-IRES-ZsGreen). The lentivirus line was generatedusing Clontech's standard protocols, and high-expressing cells wereenriched by sorting for cells expressing green fluorescent protein.BAFF(134-285) expressing HEK293F cells were incubated for 96 hoursbefore cells were pelleted and expression of the supernatant wasverified with an anti 6×His western blot (“6×His” disclosed as SEQ IDNO: 407). Supernatant purification was completed using Ni-SepharoseAffinity Chromatography as first step. Affinity purified BAFF(134-285)was polished by Size Exclusion Chromatography using Sephacryl S-400resin. 60-mer BAFF eluted as major peak that was separated from bothlarger aggregates and small molecular weight species. The molecularweight of 60-mer BAFF was confirmed by Analytical Ultracentrifugationand SEC-multi angle laser light scattering detector system.

Sequence for His-HuBAFF (134-285): (SEQ ID NO: 401)HHHHHHENLYFQGAVQGPEETVTQDCLQLIADSETPTIQKGSYTFVPWLLSFKRGSALEEKENKILVKETGYFFIYGVLYTDKTYAMGHLIQRKKVHVFGDELSLVTLFRCIQNMPETLPNNSCYSAGIAKLEEGDELQLAIPRENAQIS LDGDVTFFGALKLL

Example 7 Functional Inhibition of Antibodies to Soluble 60-Mer HumanBAFF (Refers to Tables 7 and 8)

Antibodies were assessed for the ability to neutralize soluble 60-merhuman BAFF activation of the human BAFF receptor (BAFFR). A fixedconcentration of the 60-mer (4.2 pM) BAFF was mixed in assay medium withCHO cells expressing recombinant human BAFFR and a luciferase reportersystem and stimulated for 24 hours in an incubator at 37° C., 5% CO2 inthe presence of varying doses of anti-BAFF antibodies. Luciferaseexpression was assessed at the end of the incubation to quantify thelevel of neutralization achieved. IC50 and IC90 values were determinedfrom the plots of the antibody dose titration luciferase inhibitionresults. Clinical reference antibodies (Reference 1 and Reference 2)were used as comparators.

Example 8 Functional Inhibition of Antibodies to mbBAFF (Refers toTables 7 and 8, and FIG. 2)

Antibodies were assessed for the ability to neutralize humanmembrane-bound BAFF (mbBAFF) activation of the human BAFF receptor(BAFFR). In brief, Chinese hamster ovary (CHO) cells overexpressingrecombinant human full-length BAFF sequences were produced and used as asource of cell associated (mbBAFF) BAFF. The mbBAFF-CHO were fixed inparaformaldehyde at room temperature for 1 hour with intermittentmixing. The fixed cells were washed and resuspended in complete mediumfor incubation at 37C and 5% CO₂ overnight. The next day, the fixedmbBAFF cells were mixed in assay medium at a of 1:3 ratio with CHO cellsexpressing recombinant human BAFFR and a luciferase reporter system andstimulated for 24 hours in an incubator at 37° C., 5% CO2 in thepresence of varying doses of anti-BAFF antibodies. Luciferase expressionwas assessed at the end of the incubation to quantify the level ofneutralization achieved. IC50 and IC90 values were determined from theplots of the antibody dose titration luciferase inhibition results.Clinical reference antibodies (Reference 1 and Reference 2) were used ascomparators.

BAFF can exist in three forms: membrane bound (mbBAFF), soluble trimericBAFF, and soluble 60-mer BAFF. The relative importance of the variousforms of BAFF in normal and disease physiology is not well understood.In previous studies, soluble BAFF was treated as a single entity(Manetta et al., Journal of Inflammation Research, 2014:7, 121-131). Inthe present invention, soluble trimeric and 60-mer human BAFF proteins,as well as human mbBAFF, were explicitly generated, and their polymericstatus confirmed. In functional assays, novel anti-BAFF antibodiesdescribed herein showed profiles that were different from the tworeference antibodies (Reference 1 and Reference 2) in their ability toneutralize soluble trimeric human BAFF, soluble 60-mer human BAFF andmembrane-bound human BAFF activation of the human BAFF receptor (BAFFR).

Example 9 Epitope Mapping of Antibodies

Hydrogen/Deuterium Exchange Mass Spectrometry (HXMS) was employed to mapthe epitope of IgG antibodies comprising mouse variable regions (Tables1 and 2) binding to human BAFF (subsequence amino acid positions134-285, tumor necrosis actor ligand superfamily member 13b, solubleform). This method determined the susceptibility of the amide backbonehydrogens of BAFF to exchange with D₂O. The experiment was conductedwith BAFF alone and BAFF with added antibodies (with deuterium). Regionsof the BAFF sequence showing significant protection from exchange due tobinding of antibodies were thus identified. Resolution of the method isdetermined by the peptides produced by digestion with pepsin. These BAFFderived peptides were identified by additional control experiments withunexchanged samples employing standard accurate mass and HPLC MS/MStechnologies.

Recombinant human BAFF was used (SEQ ID NO: 401). For eachprotein+antibody sample, an equimolar amount of BAFF (0.48 mg/mL) andantibody was incubated for 15 minutes at room temperature. A LEAPHDX-PAL system was used for all sample handling. Using the LEAP robotsystem (exchange plate kept at 25 degrees C., sample/quench plate keptat 4 degrees C.), 8 μL of sample was added to 80 μL of exchange buffer(10 mM NaH2PO4 in D2O, pH=7.4 or 10 mM NaH2PO4 in H2O, pH=7.4), mixed,and allowed to exchange for various times (60, 120, and 240 seconds). 80μL of this solution was then transferred to 80 μL of quench buffer (4MGuanidine-HCL, 0.5M TCEP-HCl), mixed, and kept at 4 degrees C. for 60seconds. 60 μL of this solution was then injected and flowed over apepsin column (2.1 mm×30 mm, Applied Biosystems), and flowed onto aMichrom C18 trap cartridge. The cartridge was washed with H₂O+0.1%formic acid for 2 minutes at 100 μL/min. A valve was then switched andthe cartridge eluted onto a Phenomenex Jupiter CS column, 1.0×50 mm,Sum, 300A. Mobile Phase A was water/acetonitrile/formic acid (99/1/0.1)and Mobile Phase B was acetonitrile/water/formic acid (95/5/0.1). Flowrate was 100 ul/min. Gradient was: 0 minutes (0% B), 6 minutes (40% B),7 minutes (40% B), 8 minutes (90% B), 10 minutes (90% B), 11 minutes (0%B). The LEAP system precools the mobile phase to ˜4 degrees C. MassSpectrometry is performed on a Thermo Orbitrap Velos (0900865). For theMS experiments (used to quantitate exchange with the D2O buffer), asingle scan method from 300-2000 for 14 minutes was used at resolution60,000. For the MS/MS experiments (used to ID peptides with the H2Oexchange buffer), a method with 7 scans was used for 14 minutes. Thefirst scan was a full range scan from 300-2000 at 60,000 resolution.Subsequent scans were CID scans of the 6 most intense ions from scan #1.Isolation width was 1.5 amu, collision energy was 35V, and activationtime was 30 msec.

MS/MS data was analyzed with the program Proteome Discoverer 1.3 (ThermoScientific). Briefly, the program uses the accurate molecular weight ofthe precursor ion and the fragmentation data for the product ions tomatch regions of the protein sequence. From this analysis, peptidesproduced from pepsin were identified. MS data was analyzed with thein-house program BI-SHAFT. Briefly, the list of peptic peptides, as wellas their charge state and retention time, and the protein sequence wereentered. The program then searches for data meeting the accurate masscriteria and calculates the average molecular weight of the isotopicdistribution. The data is inspected to identify errors, and where errorsoccur, manual calculations are done using Microsoft Excel whennecessary. Areas of protection are identified by comparing the controldata (protein alone) to the experimental data (protein with antibody).Regions of protection are indicative of binding.

The regions of the BAFF sequence showing significant protection fromexchange due to binding of antibodies (light/heavy chains comprising SEQID NOS: 49/67, 57/75, 41/58, 43/61, 45/63, 47/65, 51/69 and 53/71) wereidentified as amino acid residues 17 to 31 (SEQ ID NO: 403), 68 to 90(SEQ ID NO: 404), 126 to 137 (SEQ ID NO: 405) and 137 to 145 (SEQ ID NO:406).

TABLE 9 Epitope Mapping Sequences. Name Amino Acid Sequence RecombinantMAVQGPEETVTQDCLQLIADSETPTIQKGSYTFVPWLLSF HumanKRGSALEEKENKIVKETGYFFIYGQVLYTDKTYAMGHLI BAFFQRKKVHVFGDELSLVTLFRCIQNMPETLPNNSCYSAGIAKLEEGDELQLAIPRENAQISLDGDVTFFGALKLL (SEQ ID NO: 402) Amino acidIADSETPTIQKGSYT (SEQ ID NO: 403) position 17-31* Amino acidYTDKTYAMGHLIQRKKVHVFGDE position (SEQ ID NO: 404) 68-90* Amino AcidLQLAIPRENAQI (SEQ ID NO: 405) position 126-137* Amino acidISLDGDVTF (SEQ ID NO: 406) position 137-145* *N′ methionine ofrecombinant human BAFF not counted towards position number.

The invention claimed is:
 1. An anti-BAFF antibody molecule comprising alight chain variable domain with a CDR1 selected from the groupconsisting of any one of SEQ ID NO: 1, 5, 10, 13, 15, 76, 77, 78, 79,80, 249, 250, 251, 252, 253, 254, 255, 256, 257, 258, 259, 260, 261,262, 263, 264, 265, 266, 267, 268, 269, 270, 271, 272, 273, 274 and 275,a CDR2 selected from the group consisting of any one of SEQ ID NO: 2, 6,8, 11, 16, 276, 277, 278, 279, 280, 281, 282, 283, 284, 285, 286, 287,288, 289, 290, 291 and 292, and a CDR3 selected from the groupconsisting of any one of SEQ ID NO: 3, 4, 7, 9, 12, 14, 17, 293, 294,295, 296, 297, 298, 299, 300, 301, 302, 303, 304, 305, 306, 307, 308,309, 310 and 311; and a heavy chain variable domain with a CDR1 selectedfrom the group consisting of any one of SEQ ID NO: 18, 21, 23, 25, 28,31, 34, 36, 37, 81, 312, 313, 314, 315, 316, 317, 318, 319, 320, 321,322, 323, 324, 325, 326, 327, 328, 329, 330, 331, 332, 392, 333, 334,335 and 336, a CDR2 selected from the group consisting of any one of SEQID NO: 19, 24, 26, 29, 32, 35, 38, 337, 338, 339, 340, 341, 342, 343,344, 345, 346, 347, 348, 349, 350, 351, 352, 353, 354, 355, 356, 357,358, 359, 360, 361, 362, 363, 364, 365, 366 and 367, and a CDR3 selectedfrom the group consisting of any one of SEQ ID NO: 20, 22, 27, 30, 33,39, 368, 369, 370, 371, 372, 373, 374, 375, 376, 377, 378, 379, 380,381, 382, 383, 384, 385, 386, 387, 388, 389, 390 and
 391. 2. Ananti-BAFF antibody molecule comprising: (a) a light chain variabledomain comprising a CDR1 of SEQ ID NO: 1, a CDR2 of SEQ ID NO: 2 and aCDR3 of SEQ ID NO: 3, and a heavy chain variable domain comprising aCDR1 of SEQ ID NO: 18, a CDR2 of SEQ ID NO: 19 and a CDR3 of SEQ ID NO:20; or (b) a light chain variable domain comprising a CDR1 of SEQ ID NO:1, a CDR2 of SEQ ID NO: 2 and a CDR3 of SEQ ID NO: 4, and a heavy chainvariable domain comprising a CDR1 of SEQ ID NO: 21, a CDR2 of SEQ ID NO:19 and a CDR3 of SEQ ID NO: 22; or (c) a light chain variable domaincomprising a CDR1 of SEQ ID NO: 1, a CDR2 of SEQ ID NO: 2 and a CDR3 ofSEQ ID NO: 4 and a heavy chain variable domain comprising a CDR1 of SEQID NO: 23, a CDR2 of SEQ ID NO: 24 and a CDR3 of SEQ ID NO: 20; or (d) alight chain variable domain comprising a CDR1 of SEQ ID NO: 5, a CDR2 ofSEQ ID NO: 6 and a CDR3 of SEQ ID NO: 7 and a heavy chain variabledomain comprising a CDR1 of SEQ ID NO: 25, a CDR2 of SEQ ID NO: 26 and aCDR3 of SEQ ID NO: 27; or (e) a light chain variable domain comprising aCDR1 of SEQ ID NO: 5, a CDR2 of SEQ ID NO: 8 and a CDR3 of SEQ ID NO: 9and a heavy chain variable domain comprising a CDR1 of SEQ ID NO: 28, aCDR2 of SEQ ID NO: 29 and a CDR3 of SEQ ID NO: 30; or (f) a light chainvariable domain comprising a CDR1 of SEQ ID NO: 10, a CDR2 of SEQ ID NO:11 and a CDR3 of SEQ ID NO: 12 and a heavy chain variable domaincomprising a CDR1 of SEQ ID NO: 31, a CDR2 of SEQ ID NO: 32 and a CDR3of SEQ ID NO: 33; or (g) a light chain variable domain comprising a CDR1of SEQ ID NO: 13, a CDR2 of SEQ ID NO: 6 and a CDR3 of SEQ ID NO: 14 anda heavy chain variable domain comprising a CDR1 of SEQ ID NO: 34, a CDR2of SEQ ID NO: 35 and a CDR3 of SEQ ID NO: 27; or (h) a light chainvariable domain comprising a CDR1 of SEQ ID NO: 10, a CDR2 of SEQ ID NO:6 and a CDR3 of SEQ ID NO: 7 and a heavy chain variable domaincomprising a CDR1 of SEQ ID NO: 36, a CDR2 of SEQ ID NO: 26 and a CDR3of SEQ ID NO: 27; or (i) a light chain variable domain comprising a CDR1of SEQ ID NO: 15, a CDR2 of SEQ ID NO: 16 and a CDR3 of SEQ ID NO: 17and a heavy chain variable domain comprising a CDR1 of SEQ ID NO: 37, aCDR2 of SEQ ID NO: 38 and a CDR3 of SEQ ID NO: 39; or (j) a light chainvariable domain comprising a CDR1 of SEQ ID NO: 76, a CDR2 of SEQ ID NO:16 and a CDR3 of SEQ ID NO: 17 and a heavy chain variable domaincomprising a CDR1 of SEQ ID NO: 37, a CDR2 of SEQ ID NO: 38 and a CDR3of SEQ ID NO: 39; or (k) a light chain variable domain comprising a CDR1of SEQ ID NO: 77, a CDR2 of SEQ ID NO: 16 and a CDR3 of SEQ ID NO: 17and a heavy chain variable domain comprising a CDR1 of SEQ ID NO: 37, aCDR2 of SEQ ID NO: 38 and a CDR3 of SEQ ID NO: 39; or (l) a light chainvariable domain comprising a CDR1 of SEQ ID NO: 78, a CDR2 of SEQ ID NO:16 and a CDR3 of SEQ ID NO: 17 and a heavy chain variable domaincomprising a CDR1 of SEQ ID NO: 37, a CDR2 of SEQ ID NO: 38 and a CDR3of SEQ ID NO: 39; or (m) a light chain variable domain comprising a CDR1of SEQ ID NO: 79, a CDR2 of SEQ ID NO: 16 and a CDR3 of SEQ ID NO: 17and a heavy chain variable domain comprising a CDR1 of SEQ ID NO: 37, aCDR2 of SEQ ID NO: 38 and a CDR3 of SEQ ID NO: 39; or (n) a light chainvariable domain comprising a CDR1 of SEQ ID NO: 80, a CDR2 of SEQ ID NO:16 and a CDR3 of SEQ ID NO: 17 and a heavy chain variable domaincomprising a CDR1 of SEQ ID NO: 37, a CDR2 of SEQ ID NO: 38 and a CDR3of SEQ ID NO: 39; or (o) a light chain variable domain comprising a CDR1of SEQ ID NO: 5, a CDR2 of SEQ ID NO: 8 and a CDR3 of SEQ ID NO: 9 and aheavy chain variable domain comprising a CDR1 of SEQ ID NO: 81, a CDR2of SEQ ID NO: 29 and a CDR3 of SEQ ID NO: 30; or (p) a light chainvariable domain comprising a CDR1 of SEQ ID NO: 249, a CDR2 of SEQ IDNO: 276 and a CDR3 of SEQ ID NO: 293 and a heavy chain variable domaincomprising a CDR1 of SEQ ID NO: 312, a CDR2 of SEQ ID NO: 337 and a CDR3of SEQ ID NO: 368; or (q) a light chain variable domain comprising aCDR1 of SEQ ID NO: 250, a CDR2 of SEQ ID NO: 276 and a CDR3 of SEQ IDNO: 293 and a heavy chain variable domain comprising a CDR1 of SEQ IDNO: 312, a CDR2 of SEQ ID NO: 337 and a CDR3 of SEQ ID NO: 368; or (r) alight chain variable domain comprising a CDR1 of SEQ ID NO: 251, a CDR2of SEQ ID NO: 277 and a CDR3 of SEQ ID NO: 294 and a heavy chainvariable domain comprising a CDR1 of SEQ ID NO: 313, a CDR2 of SEQ IDNO: 338 and a CDR3 of SEQ ID NO: 369; or (s) a light chain variabledomain comprising a CDR1 of SEQ ID NO: 252, a CDR2 of SEQ ID NO: 278 anda CDR3 of SEQ ID NO: 295 and a heavy chain variable domain comprising aCDR1 of SEQ ID NO: 314, a CDR2 of SEQ ID NO: 339 and a CDR3 of SEQ IDNO: 370; or (t) a light chain variable domain comprising a CDR1 of SEQID NO: 253, a CDR2 of SEQ ID NO: 279 and a CDR3 of SEQ ID NO: 296 and aheavy chain variable domain comprising a CDR1 of SEQ ID NO: 315, a CDR2of SEQ ID NO: 340 and a CDR3 of SEQ ID NO: 371; or (u) a light chainvariable domain comprising a CDR1 of SEQ ID NO: 15, a CDR2 of SEQ ID NO:16 and a CDR3 of SEQ ID NO: 297 and a heavy chain variable domaincomprising a CDR1 of SEQ ID NO: 316, a CDR2 of SEQ ID NO: 341 and a CDR3of SEQ ID NO: 39; or (v) a light chain variable domain comprising a CDR1of SEQ ID NO: 254, a CDR2 of SEQ ID NO: 280 and a CDR3 of SEQ ID NO: 298and a heavy chain variable domain comprising a CDR1 of SEQ ID NO: 317, aCDR2 of SEQ ID NO: 342 and a CDR3 of SEQ ID NO: 372; or (w) a lightchain variable domain comprising a CDR1 of SEQ ID NO: 255, a CDR2 of SEQID NO: 281 and a CDR3 of SEQ ID NO: 298 and a heavy chain variabledomain comprising a CDR1 of SEQ ID NO: 317, a CDR2 of SEQ ID NO: 343 anda CDR3 of SEQ ID NO: 373; or (x) a light chain variable domaincomprising a CDR1 of SEQ ID NO: 256, a CDR2 of SEQ ID NO: 2 and a CDR3of SEQ ID NO: 299 and a heavy chain variable domain comprising a CDR1 ofSEQ ID NO: 317, a CDR2 of SEQ ID NO: 344 and a CDR3 of SEQ ID NO: 372;or (y) a light chain variable domain comprising a CDR1 of SEQ ID NO:255, a CDR2 of SEQ ID NO: 281 and a CDR3 of SEQ ID NO: 298 and a heavychain variable domain comprising a CDR1 of SEQ ID NO: 318, a CDR2 of SEQID NO: 343 and a CDR3 of SEQ ID NO: 374; or (z) a light chain variabledomain comprising a CDR1 of SEQ ID NO: 257, a CDR2 of SEQ ID NO: 282 anda CDR3 of SEQ ID NO: 300 and a heavy chain variable domain comprising aCDR1 of SEQ ID NO: 319, a CDR2 of SEQ ID NO: 345 and a CDR3 of SEQ IDNO: 375; or (aa) a light chain variable domain comprising a CDR1 of SEQID NO: 258, a CDR2 of SEQ ID NO: 283 and a CDR3 of SEQ ID NO: 301 and aheavy chain variable domain comprising a CDR1 of SEQ ID NO: 320, a CDR2of SEQ ID NO: 346 and a CDR3 of SEQ ID NO: 376; or (bb) a light chainvariable domain comprising a CDR1 of SEQ ID NO: 259, a CDR2 of SEQ IDNO: 281 and a CDR3 of SEQ ID NO: 298 and a heavy chain variable domaincomprising a CDR1 of SEQ ID NO: 317, a CDR2 of SEQ ID NO: 347 and a CDR3of SEQ ID NO: 377; or (cc) a light chain variable domain comprising aCDR1 of SEQ ID NO: 260, a CDR2 of SEQ ID NO: 284 and a CDR3 of SEQ IDNO: 294 and a heavy chain variable domain comprising a CDR1 of SEQ IDNO: 321, a CDR2 of SEQ ID NO: 348 and a CDR3 of SEQ ID NO: 378; or (dd)a light chain variable domain comprising a CDR1 of SEQ ID NO: 254, aCDR2 of SEQ ID NO: 2 and a CDR3 of SEQ ID NO: 299 and a heavy chainvariable domain comprising a CDR1 of SEQ ID NO: 322, a CDR2 of SEQ IDNO: 349 and a CDR3 of SEQ ID NO: 372; or (ee) a light chain variabledomain comprising a CDR1 of SEQ ID NO: 261, a CDR2 of SEQ ID NO: 285 anda CDR3 of SEQ ID NO: 294 and a heavy chain variable domain comprising aCDR1 of SEQ ID NO: 323, a CDR2 of SEQ ID NO: 350 and a CDR3 of SEQ IDNO: 378; or (ff) a light chain variable domain comprising a CDR1 of SEQID NO: 262, a CDR2 of SEQ ID NO: 286 and a CDR3 of SEQ ID NO: 302 and aheavy chain variable domain comprising a CDR1 of SEQ ID NO: 324, a CDR2of SEQ ID NO: 351 and a CDR3 of SEQ ID NO: 379; or (gg) a light chainvariable domain comprising a CDR1 of SEQ ID NO: 263, a CDR2 of SEQ IDNO: 6 and a CDR3 of SEQ ID NO: 303 and a heavy chain variable domaincomprising a CDR1 of SEQ ID NO: 317, a CDR2 of SEQ ID NO: 352 and a CDR3of SEQ ID NO: 380; or (hh) a light chain variable domain comprising aCDR1 of SEQ ID NO: 264, a CDR2 of SEQ ID NO: 276 and a CDR3 of SEQ IDNO: 304 and a heavy chain variable domain comprising a CDR1 of SEQ IDNO: 325, a CDR2 of SEQ ID NO: 353 and a CDR3 of SEQ ID NO: 381; or (ii)a light chain variable domain comprising a CDR1 of SEQ ID NO: 265, aCDR2 of SEQ ID NO: 287 and a CDR3 of SEQ ID NO: 305 and a heavy chainvariable domain comprising a CDR1 of SEQ ID NO: 326, a CDR2 of SEQ IDNO: 354 and a CDR3 of SEQ ID NO: 382; or (jj) a light chain variabledomain comprising a CDR1 of SEQ ID NO: 266, a CDR2 of SEQ ID NO: 287 anda CDR3 of SEQ ID NO: 306 and a heavy chain variable domain comprising aCDR1 of SEQ ID NO: 326, a CDR2 of SEQ ID NO: 355 and a CDR3 of SEQ IDNO: 383; or (kk) a light chain variable domain comprising a CDR1 of SEQID NO: 267, a CDR2 of SEQ ID NO: 285 and a CDR3 of SEQ ID NO: 294 and aheavy chain variable domain comprising a CDR1 of SEQ ID NO: 327, a CDR2of SEQ ID NO: 356 and a CDR3 of SEQ ID NO: 369; or (ll) a light chainvariable domain comprising a CDR1 of SEQ ID NO: 268, a CDR2 of SEQ IDNO: 276 and a CDR3 of SEQ ID NO: 306 and a heavy chain variable domaincomprising a CDR1 of SEQ ID NO: 328, a CDR2 of SEQ ID NO: 357 and a CDR3of SEQ ID NO: 383; or (mm) a light chain variable domain comprising aCDR1 of SEQ ID NO: 269, a CDR2 of SEQ ID NO: 288 and a CDR3 of SEQ IDNO: 304 and a heavy chain variable domain comprising a CDR1 of SEQ IDNO: 329, a CDR2 of SEQ ID NO: 358 and a CDR3 of SEQ ID NO: 384; or (nn)a light chain variable domain comprising a CDR1 of SEQ ID NO: 270, aCDR2 of SEQ ID NO: 276 and a CDR3 of SEQ ID NO: 306 and a heavy chainvariable domain comprising a CDR1 of SEQ ID NO: 330, a CDR2 of SEQ IDNO: 359 and a CDR3 of SEQ ID NO: 385; or (oo) a light chain variabledomain comprising a CDR1 of SEQ ID NO: 371, a CDR2 of SEQ ID NO: 289 anda CDR3 of SEQ ID NO: 307 and a heavy chain variable domain comprising aCDR1 of SEQ ID NO: 331, a CDR2 of SEQ ID NO: 360 and a CDR3 of SEQ IDNO: 385; or (pp) a light chain variable domain comprising a CDR1 of SEQID NO: 261, a CDR2 of SEQ ID NO: 285 and a CDR3 of SEQ ID NO: 294 and aheavy chain variable domain comprising a CDR1 of SEQ ID NO: 332, a CDR2of SEQ ID NO: 361 and a CDR3 of SEQ ID NO: 386; or (qq) a light chainvariable domain comprising a CDR1 of SEQ ID NO: 272, a CDR2 of SEQ IDNO: 289 and a CDR3 of SEQ ID NO: 307 and a heavy chain variable domaincomprising a CDR1 of SEQ ID NO: 331, a CDR2 of SEQ ID NO: 362 and a CDR3of SEQ ID NO: 385; or (rr) a light chain variable domain comprising aCDR1 of SEQ ID NO: 266, a CDR2 of SEQ ID NO: 287 and a CDR3 of SEQ IDNO: 306 and a heavy chain variable domain comprising a CDR1 of SEQ IDNO: 326, a CDR2 of SEQ ID NO: 355 and a CDR3 of SEQ ID NO: 383; or (ss)a light chain variable domain comprising a CDR1 of SEQ ID NO: 270, aCDR2 of SEQ ID NO: 276 and a CDR3 of SEQ ID NO: 306 and a heavy chainvariable domain comprising a CDR1 of SEQ ID NO: 330, a CDR2 of SEQ IDNO: 359 and a CDR3 of SEQ ID NO: 385; or (tt) a light chain variabledomain comprising a CDR1 of SEQ ID NO: 270, a CDR2 of SEQ ID NO: 276 anda CDR3 of SEQ ID NO: 306 and a heavy chain variable domain comprising aCDR1 of SEQ ID NO: 392, a CDR2 of SEQ ID NO: 363 and a CDR3 of SEQ IDNO: 387; or (uu) a light chain variable domain comprising a CDR1 of SEQID NO: 273, a CDR2 of SEQ ID NO: 276 and a CDR3 of SEQ ID NO: 308 and aheavy chain variable domain comprising a CDR1 of SEQ ID NO: 333, a CDR2of SEQ ID NO: 364 and a CDR3 of SEQ ID NO: 388; or (vv) a light chainvariable domain comprising a CDR1 of SEQ ID NO: 274, a CDR2 of SEQ IDNO: 290 and a CDR3 of SEQ ID NO: 309 and a heavy chain variable domaincomprising a CDR1 of SEQ ID NO: 334, a CDR2 of SEQ ID NO: 365 and a CDR3of SEQ ID NO: 389; or (ww) a light chain variable domain comprising aCDR1 of SEQ ID NO: 275, a CDR2 of SEQ ID NO: 291 and a CDR3 of SEQ IDNO: 310 and a heavy chain variable domain comprising a CDR1 of SEQ IDNO: 335, a CDR2 of SEQ ID NO: 366 and a CDR3 of SEQ ID NO: 390; or (xx)a light chain variable domain comprising a CDR1 of SEQ ID NO: 258, aCDR2 of SEQ ID NO: 292 and a CDR3 of SEQ ID NO: 311 and a heavy chainvariable domain comprising a CDR1 of SEQ ID NO: 336, a CDR2 of SEQ IDNO: 367 and a CDR3 of SEQ ID NO:
 391. 3. An anti-BAFF antibody moleculecomprising a light chain variable domain of any one of SEQ ID NOS:82-97, and a heavy chain variable domain of any one of SEQ ID NOS:100-115.
 4. The anti-BAFF antibody molecule of claim 3, wherein acombination of light chain variable domain and heavy chain variabledomain comprises SEQ ID NOS: 82/101, 88/101, 94/112 or 93/114.
 5. Theanti-BAFF antibody molecule of claim 2, wherein the anti-BAFF antibodymolecule neutralizes all three forms of human BAFF, the forms of whichinclude membrane bound (mbBAFF), soluble trimeric BAFF, and soluble60-mer BAFF.
 6. The anti-BAFF antibody molecule of claim 2, wherein theanti-BAFF antibody molecule neutralizes human soluble trimeric BAFF. 7.The anti-BAFF antibody molecule of claim 2, wherein the anti-BAFFantibody molecule neutralizes human membrane bound BAFF.
 8. Theanti-BAFF antibody molecule of claim 2, wherein the anti-BAFF antibodymolecule neutralizes human soluble 60-mer BAFF.
 9. An anti-BAFF antibodymolecule comprising: a) a humanized light chain variable domaincomprising the CDRs of SEQ ID NO: 76, 16 and 17 and framework regionshaving an amino acid sequence at least 90% identical to the amino acidsequence of the framework regions of the variable domain light chainamino acid sequence of SEQ ID NO: 82; and b) a humanized heavy chainvariable domain comprising the CDRs of SEQ ID NO:37, 38 and 39 andframework regions having an amino acid sequence at least 90% identicalto the amino acid sequence of the framework regions of the variabledomain heavy chain amino acid sequence of SEQ ID NO:
 101. 10. Ananti-BAFF antibody molecule comprising: a) a humanized light chainvariable domain comprising the CDRs of SEQ ID NO: 15, 16 and 17 andframework regions having an amino acid sequence at least 90% identicalto the amino acid sequence of the framework regions of the variabledomain light chain amino acid sequence of SEQ ID NO: 88; and b) ahumanized heavy chain variable domain comprising the CDRs of SEQ IDNO:37, 38 and 39 and framework regions having an amino acid sequence atleast 90% identical to the amino acid sequence of the framework regionsof the variable domain heavy chain amino acid sequence of SEQ ID NO:101.
 11. An anti-BAFF antibody molecule comprising: a) a humanized lightchain variable domain comprising the CDRs of SEQ ID NO: 5, 8 and 9 andframework regions having an amino acid sequence at least 90% identicalto the amino acid sequence of the framework regions of the variabledomain light chain amino acid sequence of SEQ ID NO: 94; and b) ahumanized heavy chain variable domain comprising the CDRs of SEQ ID NO:81, 29 and 30 and framework regions having an amino acid sequence atleast 90% identical to the amino acid sequence of the framework regionsof the variable domain heavy chain amino acid sequence of SEQ ID NO:112.
 12. An anti-BAFF antibody molecule comprising: a) a humanized lightchain variable domain comprising the CDRs of SEQ ID NO: 5, 8 and 9 andframework regions having an amino acid sequence at least 90% identicalto the amino acid sequence of the framework regions of the variabledomain light chain amino acid sequence of SEQ ID NO: 93; and b) ahumanized heavy chain variable domain comprising the CDRs of SEQ ID NO:81, 29 and 30 and framework regions having an amino acid sequence atleast 90% identical to the amino acid sequence of the framework regionsof the variable domain heavy chain amino acid sequence of SEQ ID NO:114.
 13. An anti-BAFF antibody molecule according to claim 1, whereinthe antibody is a monoclonal antibody.
 14. The anti-BAFF antibodymolecule according to claim 13, wherein the monoclonal antibody is ahumanized monoclonal antibody.
 15. A pharmaceutical compositioncomprising an anti-BAFF antibody molecule according to claim 1 and apharmaceutically acceptable carrier.
 16. A method for treating a subjecthaving a BAFF-associated disorder comprising administering to thesubject an anti-BAFF antibody molecule according to claim 1, or apharmaceutical composition comprising an anti-BAFF antibody moleculeaccording to claim 1 and a pharmaceutically acceptable carrier, whereinthe anti-BAFF antibody molecule binds to human BAFF.
 17. A method fortreating an inflammatory disease, an autoimmune disease, a respiratorydisease, a metabolic disorder or cancer comprising administering to asubject in need thereof an effective amount of an anti-BAFF antibodymolecule according to claim 1, or a pharmaceutical compositioncomprising an anti-BAFF antibody molecule according to claim 1 and apharmaceutically acceptable carrier.
 18. The method according to claim17, wherein the disease is systemic lupus erythematosus, lupus nephritisor rheumatoid arthritis.
 19. A method for inhibiting the binding of BAFFto one or more BAFF receptors on a mammalian cell, wherein the BAFFreceptor is BAFF-R (BR3), TACI (transmembrane activator and calciummodulator and cyclophilin ligand interactor) and/or BCMA (B-cellmaturation antigen), comprising administering to the cell an anti-BAFFantibody molecule according to claim 1, wherein signaling mediated bythe BAFF receptor is inhibited.
 20. An isolated polynucleotidecomprising a sequence encoding a light chain variable region of any oneof SEQ ID NOS: 82-97, or a heavy chain variable region of any one of SEQID NOS: 100-115.
 21. The isolated polynucleotide of claim 20, whereinthe light chain variable region is SEQ ID NO: 234 and the heavy chainvariable region is SEQ ID NO: 396, the light chain variable region isSEQ ID NO: 393 and the heavy chain variable region is SEQ ID NO: 396,the light chain variable region is SEQ ID NO: 395 and the heavy chainvariable region is SEQ ID NO: 397 or the light chain variable region isSEQ ID NO: 394 and the heavy chain variable region is SEQ ID NO: 398.